CN101925595A

CN101925595A - Phthalazinone derivatives

Info

Publication number: CN101925595A
Application number: CN2009801027174A
Authority: CN
Inventors: 基思·艾伦·米尼尔; 穆罕默德·哈什姆·贾瓦齐; 西尔维·戈麦斯; 马克·杰夫瑞·休默索恩; 卡洛斯·费南德斯·莱恩斯; 尼尔·莫里森·巴尔·马丁; 戴维·艾伦·拉奇; 克雷格·安东尼·罗伯茨; 凯文·布雷德斯
Original assignee: AstraZeneca AB
Current assignee: AstraZeneca AB
Priority date: 2008-01-23
Filing date: 2009-01-23
Publication date: 2010-12-22
Also published as: KR20100116187A; WO2009093032A1; MX2010006364A; AR070221A1; CL2009000133A1; NI201000120A; ECSP10010349A; EA201070523A1; TW200936577A; ZA201003752B; US8129380B2; CO6290715A2; AU2009207498A1; EP2231638A1; BRPI0906436A2; PE20091335A1; NZ585982A; US20120129855A1; JP2011510056A; SV2010003625A

Abstract

A compound of the formula (I): wherein: A and B together represent an optionally substituted, fused aromatic ring; X and Y are selected from CH and CH, CF and CH, CH and CF and N and CH respectively; Rc is selected from H, C1.4 alkyl; and R1 is selected from C1-7 alkyI, C3-20 heterocydyl and C5-20 aryl, which groups are optionally substituted; or Rc and R1 together with the carbon and oxygen atoms to which they are attached form a spiro-C5-7 oxygen-containing heterocyclic group, which is optionally substituted or fused to a C5-7 aromatic ring. These compounds are useful in the preparation of a medicament for inhibiting the activity of PARP for the treatment of : vascular disease; septic shock;ischaemic injury;neurotoxicity;haemorraghic shock;viral infection; or for use as an adjunct in cancer therapy or for potentiating tumour cells for treatment with ionizing radiation or chemotherapeutic agents.

Description

Phthalazinone derivatives

The present invention relates to phthalazinone derivatives and they purposes as medicine.Particularly, the present invention relates to the poly-active purposes of (ADP-ribose) polysaccharase-1 of these compound inhibitory enzymes, this enzyme is also referred to as poly-(ADP-ribose) synthetic enzyme and poly-ADP-ribosyltransferase is commonly called PARP-1.

Mammalian enzyme PARP-1 (the Multidomain protein of a kind of 113-kDa) conducts relevant (D ' Amours in conjunction with the ability of dna single chain or double-strand break with the signal of dna damage with quick by its identification, Deng, Biochem.J., 342,249-268 (1999)).

Poly-(ADP-ribose) polysaccharase family comprises about 18 kinds of protein now, and they are in the homology that all shows certain level aspect its catalyst structure domain, but their cell function difference (Ame, etc., Bioessays., 26 (8), 882-893 (2004)).In this family, PARP-1 (founder) and PARP-2 are only up to now by the enzyme that the DNA splitting of chain activates catalytic activity takes place, and this makes them very unique in family.

Present known PARP-1 participates in various DNA correlation functions, comprise that gene amplification, cell fission, differentiation, apoptosis, the excision of DNA base are repaired and to the effect (d ' Adda di Fagagna of telomere length and chromosome stability, Deng, Nature Gen., 23 (1), 76-80 (1999)).

To PARP-1 regulate that DNA repairs and the Study on Mechanism of other processes verified in nucleus its importance (Althaus in the formation of poly-(ADP-ribose) chain, F.R. and Richter, C., ADP-Ribosylation of Proteins:Enzymology and Biological Significance, Springer-Verlag, Berlin (1987)).Utilize NAD with DNA bonded, activatory PARP-1 ⁺Go up synthetic poly-(ADP-ribose) (Rhun, etc., Biochem.Biophys.Res.Commun., 245,1-10 (1998)) in various nuclear target protein matter (comprising topoisomerase, histone and PARP self).

Poly-(ADP-ribosyl) changed also relevant with vicious transformation.For example, fibroblastic nucleus Nei Genggao that the PARP-1 activity transforms at isolating SV40-, and leukemia and colon cancer cell all show normal white corpuscle and the higher enzymic activity (Miwa of mucous membrane of colon than equivalent, Deng, Arch.Biochem.Biophys., 181,313-321 (1977); Burzio, etc., Proc.Soc.Exp.Biol.Med., 149,933-938 (1975); And Hirai, etc., Cancer Res., 43,3441-3446 (1983)).Verifiedly recently compare with the benign prostate cell, the remarkable rising of active PARP in the malignant prostate tumour (mainly being PARP-1) level is relevant with higher levels of genetic instability, and (McNealy is etc., Anticancer Res., 23,1473-1478 (2003)).

Many low-molecular-weight PARP-1 inhibitor have been used for explaination poly-(ADP-ribosyl) and have changed the functional effect of repairing at DNA.In the cell of handling with alkylating agent, the inhibition of PARP cause the remarkable increase that DNA-splitting of chain and cell kill (Durkacz, etc., Nature, 283,593-596 (1980); Berger, N.A., Radiation Research, 101,4-14 (1985)).

Afterwards, this inhibitor be proved by the reparation that suppresses PLD strengthen the effect that radiation responds (Ben-Hur, etc., British Journal of Cancer, 49 (Suppl.VI), 34-42 (1984); Schlicker, etc., Int.J.Radiat.Bioi., 75,91-100 (1999)).It is reported, the PARP inhibitor in the hypoxic tumor cells of radiation sensitization effectively (US 5,032,617; US 5,215,738 and US 5,041,653).In some tumor cell line, the active Chemical Inhibition of PARP-1 (with PARP-2) also with remarkable sensitization relevant (Chalmers, Clin.Oncol., 16 (1), 29-39 (2004)) to the radiation of unusual low dosage.

In addition, the animal that PARP-1 knocks out (PARP-/-) alkylating agent and γ-radiation response are shown genomic instability (Wang, etc., Genes Dev., 9,509-520 (1995); Menissier de Murcia, etc., Proc.Natl.Acad.Sci.USA, 94,7303-7307 (1997)).Nearest data show that PARP-1 and PARP-2 are keeping having overlapping and the nonredundancy function on the genome stability, this makes and they all becomes interesting target spot (Menissier de Murcia is etc., EMBO.J., 22 (9), 2255-2263 (2003)).

Recently report also that the PARP restraining effect has blood vessel formation against function.The dose-dependently of having reported VEGF reduces and HUVECS neutral and alkali fibroblast growth factor (bFGF) inductive propagation, migration and pipe formation (Rajesh, etc., Biochem.Biophys.Res.Comm., 350,1056-1062 (2006)).

In some vascular disease, septic shock, ischemia injury and neurotoxicity, also confirmed PARP-1 effect (Cantoni, etc., Biochim.Biophys.Acta, 1014,1-7 (1989); Szabo, etc., J.Clin.Invest., 100,723-735 (1997)).Causing subsequently the oxyradical dna damage of the DNA splitting of chain discerned by PARP-1 is the main reason of this morbid state, shown in the research of PARP-1 inhibitor (Cosi, etc., J.Neurosci.Res., 39,38-46 (1994); Said, etc., Proc.Natl.Acad.Sci.U.S.A., 93,4688-4692 (1996)).Recently, confirmed PARP in the morbidity of hemorrhagic shock, work (Liaudet, etc., Proc.Natl.Acad.Sci.U.S.A., 97 (3), 10203-10208 (2000)).

Also confirm and to stop the effective retroviral infection of mammalian cell by suppressing the PARP-1 activity.Shown that (Gaken, etc., J.Virology, 70 (6), 3992-4000 (1996)) takes place this restraining effect that recombinant retroviral vector infects in various cell type.Therefore, the inhibitor of PARP-1 has been developed for antiviral therapy and cancer therapy (WO 91/18591).

In addition, infer that the PARP-1 restraining effect can delay the beginning (Rattan and Clark, Biochem.Biophys.Res.Comm., 201 (2), 665-672 (1994)) of aging characteristics among the human fibroblast.This may be relevant with PARP role in control telomere function (d ' Adda di Fagagna, etc., Nature Gen., 23 (1), 76-80 (1999)).

Also think PARP inhibitor and inflammatory bowel (Szabo C., Role of Poly (ADP-Ribose) Polymerase Activation in the Pathogenesis of Shock and Inflammation, In PARP as a Therapeutic Target; Ed J.Zhang, 2002, CRC Press; 169-204), ulcerative colitis (Zingarelli, B is etc., Immunology, 113 (4), 509-517 (2004)) and Crohn disease (Jijon, H.B., etc., Am.J.Physiol.Gastrointest.Liver Physiol., 279, G641-G651 (2000)) treatment is relevant.

1 (the 2H)-phthalazone compound of (WO 2004/080976) class as the PARP inhibitor described before contrivers more of the present invention.Described compound has general formula:

Wherein:

A and B represent the optional fused aromatic rings that replaces together;

X can be NR ^XOr CR ^XR ^Y

If X=NR ^XIf then n is 1 or 2 and X=CR ^XR ^YThen n is 1;

R ^XBe selected from H, the optional C that replaces _1-20Alkyl, C _5-20Aryl, C _3-20Heterocyclic radical, amido, thio acylamino, sulfonamido, ester, aryl and alkylsulfonyl;

R ^YBe selected from H, hydroxyl, amino;

Perhaps R ^XAnd R ^YForm spiral shell C together _3-7Cycloalkyl or heterocyclic radical;

R ^C1And R ^C2All be hydrogen, perhaps working as X is CR ^XR ^YThe time, R ^C1, R ^C2, R ^XAnd R ^YCan form the optional fused aromatic rings that replaces with the carbon atom that they connected; And

R ¹Be selected from H and halogen.

The inventor has now found that X is CHR ^XAnd R ^XFor the compound of ether demonstrates surprising lifting in the active inhibition level of PARP and/or radiotherapy and various chemotherapy aspect the enhancement of tumour cell treatment horizontal.These compounds also demonstrate the solvability of raising and more difficultly flow out from cell, thereby have improved bioavailability.

Therefore, a first aspect of the present invention provides the compound of formula (I):

Wherein:

A and B represent the optional fused aromatic rings that replaces together;

X and Y are selected from CH and CH, CF and CH, CH and CF and N and CH respectively;

R ^CBe selected from H, C _1-4Alkyl; And

R ¹Be selected from C _1-7Alkyl, C _3-20Heterocyclic radical and C _5-20Aryl, described group are optional the replacements; Perhaps

R ^CAnd R ¹Form spiral shell C with carbon atom and Sauerstoffatom that they connected _5-7Oxygen-containing heterocycle, described heterocyclic radical are optional replace or and C _5-7The aromatic ring condensed.

Therefore, work as R ^CWhen being CH for H and Y, described compound has the structure of formula (Ia):

A second aspect of the present invention provides the compound that comprises first aspect and the pharmaceutical composition of pharmaceutical acceptable carrier or thinner.

The compound that a third aspect of the present invention provides first aspect is to the purposes in the methods of treatment of human body and animal body.

Fourth aspect present invention provides as the purposes of the defined compound of first aspect present invention in medication preparation, and described medicine is used for:

(a) prevent to gather (ADP-ribose) chain formation by the activity that suppresses cell PARP (PARP-1 and/or PARP-2);

(b) treatment: vascular disease; Septic shock; The cerebrovascular and cardiovascular ischemia injury; The cerebrovascular and cardiovascular reperfusion injury; Neurotoxicity comprises apoplexy and parkinsonian acute and chronic treatment; Hemorrhagic shock; Inflammatory diseases, for example sacroiliitis, inflammatory bowel, ulcerative colitis and Crohn disease; Multiple sclerosis; The diabetes secondary effect; And the Cytotoxic acute treatment behind the operation on vessels of heart or by suppressing the active disease of improving of PARP;

(c) in cancer therapy, use or be used to strengthen the therapeutic action of ionizing radiation or chemotherapeutic tumour cell as auxiliary.

Particularly, the compound of first aspect present invention definition can be used from anticancer combination therapy (perhaps as auxiliary) with alkylating agent and with topoisomerase-1 inhibitor one, described alkylating agent is methyl mesylate (MMS), Temozolomide and Dacarbazine (DTIC) for example, and described topoisomerase-1 inhibitor for example Hycamtin, Rinotecan, rubitecan, exatecan, lurtotecan, gefitinib, difluoro replaces health (high camptothecin); And the Fei Silan that 7-replaces is for health (non-silatecan) class; 7-silyl camptothecin BNP 1350; With non-camptothecine topoisomerase-I inhibitor, for example indolocarbazole class, and topoisomerase-I and II double inhibitor, for example phenonaphthazine class, XR 11576/MLN 576 and benzo pyrido indoles.This combination can for example provide as iv formulation or by Orally administered according to the preferred application process of certain drug.

Other aspects of the present invention provide by suppressing the treatment of diseases that PARP is alleviated, comprise the compound that in the first aspect of experimenter's administering therapeutic significant quantity that needs are treated, defines, preferably use with the form of pharmaceutical composition, and provide treatment for cancer, comprise that the experimenter to the needs treatment makes up compound and radiotherapy (ionizing rays) or the chemotherapeutic that defines in the first aspect of administering therapeutic significant quantity, the compound that defines in the described first aspect is preferably with the form of pharmaceutical composition, with radiotherapy or chemotherapeutic simultaneously or successively use.

In other aspects of the present invention, described compound can be used for preparing the medicine of the cancer for the treatment of homologous recombination (HR) dependent DNA double-strand break (DSB) repairing activity disappearance or be used for the treatment of the cancer patients who suffers from HR dependent DNA DSB repairing activity disappearance, comprises the described compound to described patient's administering therapeutic significant quantity.

HR dependent DNA DSB reparation approach by the double-strand break (DSB) in the homology mechanism DNA plerosis to form successive DNA spiral (K.K.Khanna and S.P.Jackson, Nat.Genet.27 (3): 247-254 (2001)) again.The component that HR dependent DNA DSB repairs approach includes but not limited to ATM (NM_000051), RAD51 (NM_002875), RAD51L1 (NM_002877), RAD51C (NM_002876), RAD51L3 (NM_002878), DMC1 (NM_007068), XRCC2 (NM_005431), XRCC3 (NM_005432), RAD52 (NM_002879), RAD54L (NM_003579), RAD54B (NM_012415), BRCA1 (NM_007295), BRCA2 (NM_000059), RAD50 (NM_005732), MRE11A (NM_005590) and NBS1 (NM_002485).Repair other relevant protein of approach with HR dependent DNA DSB and comprise regulatory factor, for example EMSY (Hughes-Davies, etc., Cell, 115, pp523-535).In " Wood, etc., Science, 291,1284-1289 (2001) ", also the HR component is described.

The cancer of HR dependent DNA DSB reparation disappearance may comprise one or more cancer cells or be made up of one or more cancer cells, with respect to normal cell, they reduce or forfeiture by the ability of this approach DNA plerosis DSB, and promptly the activity of HR dependent DNA DSB reparation approach may reduce or lose in one or more cancer cells.

In suffering from one or more cancer cells of individuality of cancer that HR dependent DNA DSB repairs disappearance, HR dependent DNA DSB repairs the activity of one or more components of approach and may lose.The component that HR dependent DNA DSB repairs approach has been carried out abundant sign (referring to for example Wood, etc., Science, 291,1284-1289 (2001)) in the art, and comprises component listed above.

In some preferred embodiments, described cancer cells may have BRCA1 and/or BRCA2 disappearance phenotype, and promptly BRCA1 and/or BRCA2 are active in cancer cells reduces or forfeiture.Cancer cells with this phenotype may be BRCA1 and/or BRCA2 disappearance, promptly the expression of BRCA1 and/or BRCA2 and/or activity may reduce or lose in cancer cells, for example pass through the sudden change or the polymorphism of coding nucleic acid, or for example encode by the gene of the coding and regulating factor that (Hughes-Davies is etc., Cell for amplification, sudden change or the polymorphism of EMSY gene of BRCA2 regulatory factor, 115,523-535), perhaps by epigenetic mechanism, for example gene promoter methylation.

BRCA1 and BRCA2 are known tumor suppressor genes, and their wild-type allele usually lacks (Jasin M., Oncogene, 21 (58), 8981-93 (2002) in heterozygote carrier's tumour; Tutt, etc., Trends Mol Med., 8 (12), 571-6, (2002)).BRCA1 and/or BRCA2 sudden change have been carried out abundant sign (Radice, P.J., Exp.Clin.Cancer Res., 21 (3Suppl), 9-12 (2002)) in the art with getting in touch of mammary cancer.Also the amplification of the EMSY gene of known coded BRCA2 binding factor is also relevant with mammary cancer and ovarian cancer.

The carrier of BRCA1 and/or BRCA2 sudden change also be in have ovarian cancer, in the excessive risk of prostate cancer and carcinoma of the pancreas.

In some preferred embodiments, described individuality is heterozygosis with regard to one or more variations of BRCA1 and/or BRCA2 or its regulatory factor with regard to sudden change and polymorphism.The detection of BRCA1 and BRCA2 variation is well known in the art, in for example EP 699 754, EP 705 903, " Neuhausen; S.L. and Ostrander, E.A., Genet.Test; 1; 75-83 (1992) ", " Janatova M., etc., Neoplasma; 50 (4), 246-50 (2003) ", be described.Mensuration to BRCA2 binding factor EMSY amplification in " Hughes-Davies, etc., Cell, 115,523-535 " is described.

Can detect with protein level with nucleic acid level or by the existence that detects variation (i.e. sudden change or allelotrope modification) polypeptide by the existence that detects the variant nucleic acid sequence with the sudden change and the polymorphism of related to cancer.

Accompanying drawing

Fig. 1 is the powder X-ray RD figure of a crystal formation of compound of the present invention;

Fig. 2 is the DSC curve of the compound crystal formation of shown in Figure 1ization;

Fig. 3 is the powder X-ray RD figure of another crystal formation of compound shown in Figure 1;

Fig. 4 is the DSC curve of compound crystal formation shown in Figure 3;

Fig. 5 is the powder X-ray RD figure of the crystal formation of another compound of the present invention;

Fig. 6 is the DSC curve of the crystal formation of compound shown in Figure 5.

Definition

Term used herein " aromatic ring " refers to the ring-type aromatic structure at conventional meaning,, has the circulus of delocalizedπelectron track that is.

Condense with main nuclear, namely can have the aromatic ring (causing for example naphthyl or anthryl) that further condenses by-aromatic ring that A-B-forms. Described aromatic ring can only contain carbon atom, perhaps can contain carbon atom and one or more hetero atom, and described hetero atom includes but not limited to nitrogen-atoms, oxygen atom and sulphur atom. Described aromatic ring preferably has 5 or 6 annular atomses.

Described aromatic ring can be optional the replacement. If itself contains aryl substituting group, do not think that then this aryl is the part of its aryl that connects. For example, think that in this article biphenyl is the phenyl (aryl that only contains an aromatic ring) of phenyl substituted. Similarly, think that benzyl phenyl is the phenyl (aryl that only contains an aromatic ring) that benzyl replaces.

In one group of preferred embodiment, described aryl comprises an aromatic ring, and it has 5 or 6 annular atomses, and described annular atoms is selected from carbon atom, nitrogen-atoms, oxygen atom and sulphur atom, and described ring is optional the replacement. The example of these groups includes, but are not limited to benzene, pyrazine, pyrroles, thiazole, isoxazole with oxazole. Think that also the 2-pyrones is aromatic ring, but not preferred.

If described aromatic ring has 6 atoms, then at least 4 in the annular atoms or even 5 or all be carbon atom. Other annular atomses are selected from nitrogen-atoms, oxygen atom and sulphur atom, wherein preferred nitrogen atom and oxygen atom. Suitable group comprises following ring: do not have heteroatomic ring (benzene); Ring (pyridine) with 1 azo-cycle atom; Ring (pyrazine, pyrimidine and pyridazine) with 2 azo-cycle atoms; Ring (pyrones) with 1 oxygen annular atoms; With the ring (oxazine with 1 oxygen annular atoms and 1 azo-cycle atom).

If described aromatic ring has 5 annular atomses, then preferably at least 3 annular atomses are carbon atom. All the other annular atomses are selected from nitrogen-atoms, oxygen atom and sulphur atom. Suitable ring comprises following ring: the ring (pyrroles) with 1 azo-cycle atom; Ring (imidazoles, pyrazoles) with 2 azo-cycle atoms; Ring (furans) with 1 oxygen annular atoms; Ring (thiophene) with 1 sulphur annular atoms; Ring (isothiazole, thiazole) with 1 azo-cycle atom and 1 sulphur annular atoms; With the ring (isoxazole, oxazole with 1 azo-cycle atom and 1 oxygen annular atoms).

Described aromatic ring can have one or more substituting groups at arbitrary available ring position. These substituting groups are selected from halogen, nitro, hydroxyl, ether, sulfydryl, thioether, amino, C_1-7Alkyl, C_3-20Heterocyclic radical and C_5-20Aryl. Described aromatic ring can also have one or more substituting groups that form together ring. Particularly, these substituting groups can have formula-(CH₂) _m-or-O-(CH₂) _p-O-, wherein m is 2,3,4 or 5, and p is 1,2 or 3.

Alkyl: term used herein " alkyl " relates to from the carbon atom of the hydrocarbon compound with 1-20 carbon atom removes hydrogen atom and the univalent perssad (except as otherwise noted) that obtains, it can be aliphatic or alicyclic, and it can be saturated or unsaturated (for example part is unsaturated, fully unsaturated). Therefore, term " alkyl " comprises hereinafter described subclass thiazolinyl, alkynyl, cycloalkyl, cycloalkenyl group, cycloalkynyl radical etc.

In the alkyl situation, prefix (for example, C_1-4、C _1-7、C _1-20、C _2-7、C _3-7Deng) scope of expression carbon number or carbon number. For example, term " C used herein_1-4Alkyl " relate to the alkyl with 1-4 carbon atom. The example of alkyl comprises C_1-4Alkyl (" low alkyl group "), C_1-7Alkyl and C_1-20Alkyl. Notice that first prefix can change according to other restrictions; For example, for unsaturated alkyl, first prefix must be 2 at least; For cyclic alkyl, first prefix must be 3 at least; Etc..

The example of (unsubstituted) saturated alkyl includes but not limited to methyl (C₁), ethyl (C₂), propyl group (C₃), butyl (C₄), amyl group (C₅), hexyl (C₆), heptyl (C₇), octyl group (C₈), nonyl (C₉), decyl (C₁₀), undecyl (C₁₁), dodecyl (C₁₂), tridecyl (C₁₃), myristyl (C₁₄), pentadecyl (C₁₅) and eicosyl (C₂₀)。

The example of (unsubstituted) straight chain saturated alkyl includes but not limited to methyl (C₁), ethyl (C₂), n-pro-pyl (C₃), normal-butyl (C₄), n-pentyl (amyl group) (C₅), n-hexyl (C₆) and n-heptyl (C₇)。

The example of (unsubstituted) saturated branched alkyl comprises isopropyl (C₃), isobutyl group (C₄), sec-butyl (C₄), the tert-butyl group (C₄), isopentyl (C₅) and neopentyl (C₅)。

Thiazolinyl: term used herein " thiazolinyl " relates to the alkyl with one or more carbon-to-carbon double bonds. The example of thiazolinyl comprises C_2-4Thiazolinyl, C_2-7Thiazolinyl, C_2-20Thiazolinyl.

The example of (unsubstituted) unsaturated thiazolinyl includes but not limited to vinyl (CH=CH₂), 1-acrylic (CH=CH-CH₃), 2-acrylic (pi-allyl ,-CH-CH=CH₂), isopropenyl (1-methyl ethylene ,-C (CH₃)＝CH ₂), cyclobutenyl (C₄), pentenyl (C₅) and hexenyl (C₆)。

Alkynyl: term used herein " alkynyl " relates to the alkyl with one or more carbon-to-carbon three keys. The example of alkynyl comprises C_2-4Alkynyl, C_2-7Alkynyl, C_2-20Alkynyl.

The example of (unsubstituted) unsaturated alkynyl includes but not limited to acetenyl (C ≡ CH) and 2-propynyl (propargyl ,-CH₂-C≡CH)。

Cycloalkyl: it also is the alkyl of cyclic group that term used herein " cycloalkyl " relates to; That is to say, remove the univalent perssad that hydrogen atom obtains from the alicyclic annular atoms of carbocyclic compound carbocyclic ring, carbocyclic ring can be saturated or undersaturated (for example, part is undersaturated, fully undersaturated), this group has 3-20 carbon atom (except as otherwise noted), comprises 3-20 annular atoms. Therefore, term " cycloalkyl " comprises subclass cycloalkenyl group and cycloalkynyl radical. Preferably, each ring has 3-7 annular atoms. The example of cycloalkyl comprises C_3-20Cycloalkyl, C_3-15Cycloalkyl, C_3-10Cycloalkyl, C_3-7Cycloalkyl.

The example of cycloalkyl includes but not limited to the group of deriving from following compounds:

Saturated mono cyclic hydrocar-bons compound:

Cyclopropane (C₃), cyclobutane (C₄), pentamethylene (C₅), cyclohexane (C₆), cycloheptane (C₇), methyl cyclopropane (C₄), dimethylcyclopropane (C₅), methyl cyclobutane (C₅), dimethyl cyclobutane (C₆), methyl cyclopentane (C₆), dimethylcyclopentane (C₇), hexahydrotoluene (C₇), dimethyl cyclohexane (C₈), terpane (C₁₀)；

The unsaturated monocyclic hydrocarbon compounds:

Cyclopropylene (C₃), cyclobutane (C₄), cyclopentene (C₅), cyclohexene (C₆), methyl cyclopropene (C₄), dimethyl cyclopropylene (C₅), methyl cyclobutane (C₅), dimethyl cyclobutane (C₆), methyl cyclopentene (C₆), dimethylcyclopentene (C₇), methylcyclohexene (C₇), dimethyl cyclohexene (C₈)；

Saturated polycyclic hydrocarbon compound:

Thujane (C₁₀), carane (C₁₀), pinane (C₁₀), camphane (C₁₀), norcarane (C₇), norpinane (C₇), norcamphane (C₇), adamantane (C₁₀), naphthalane (decahydronaphthalene) (C₁₀)；

Unsaturated polycyclic hydrocarbon compound:

Amphene (C₁₀), citrene (C₁₀), firpene (C₁₀)；

Polycyclic hydrocarbon compound with aromatic ring:

Indenes (C ₉), indane (for example, 2,3-dihydro-1H-indenes) (C ₉), naphthane (1,2,3, the 4-naphthane) (C ₁₀), acenaphthene (C ₁₂), fluorenes (C ₁₃), non-that alkene (C ₁₃), vinegar phenanthrene (C ₁₅), aceanthrene (C ₁₆), cholanthrene (C ₂₀).

Heterocyclic radical: term used herein " heterocyclic radical " relates to from the annular atoms of heterogeneous ring compound removes the univalent perssad that hydrogen atom obtains, and this group has 3-20 annular atoms (except as otherwise noted), and wherein 1-10 is ring hetero atom.Preferably, each ring has 3-7 annular atoms, and wherein 1-4 is ring hetero atom.

In this case, prefix (for example, C _3-20, C _3-7, C _5-6Deng) be meant the annular atoms number, or the scope of annular atoms number, no matter be carbon atom or heteroatoms.For example, term " C used herein _5-6Heterocyclic radical " relate to heterocyclic radical with 5 or 6 annular atomses.The example of heterocyclic radical comprises C _3-20Heterocyclic radical, C _5-20Heterocyclic radical, C _3-15Heterocyclic radical, C _5-15Heterocyclic radical, C _3-12Heterocyclic radical, C _5-12Heterocyclic radical, C _3-10Heterocyclic radical, C _5-10Heterocyclic radical, C _3-7Heterocyclic radical, C _5-7Heterocyclic radical and C _5-6Heterocyclic radical.

The example of monocyclic heterocycles base includes but not limited to from following compounds deutero-group:

N1: aziridine (C ₃), azetidine (C ₄), tetramethyleneimine (Pyrrolidine) (C ₅), pyrroline (for example, 3-pyrroline, 2,5-pyrrolin) (C ₅), 2H-pyrroles or 3H-pyrroles's (different pyrroles, different azoles) (C ₅), piperidines (C ₆), dihydropyridine (C ₆), tetrahydropyridine (C ₆), azepines (C ₇);

O ₁: oxyethane (C ₃), trimethylene oxide (C ₄), tetrahydrofuran (tetrahydrofuran (THF)) (C ₅), oxole (dihydrofuran) (C ₅), oxane (tetrahydropyrans) (C ₆), dihydropyrane (C ₆), pyrans (C ₆), oxepin (C ₇);

S1: thiirane (C ₃), Thietane (C ₄), thiacyclopentane (tetramethylene sulfide) (C ₅), thia hexanaphthene (tetrahydric thiapyran) (C ₆), thia suberane (C ₇);

O2: dioxolane (C ₅), diox (C ₆) and Dioxepane (C ₇);

O ₃: trioxane (C ₆);

N ₂: imidazolidine (C ₅), pyrazolidine (diazolidine) (C ₅), tetrahydroglyoxaline (C ₅), pyrazoline (pyrazoline) (C ₅), piperazine (C ₆);

N ₁O ₁: Si Qing oxazole (C ₅), dihydro-oxazole (C ₅), tetrahydrochysene isoxazole (C ₅), dihydro-isoxazole (C ₅), morpholine (C ₆), Si Qing oxazine (C ₆), Er Qing oxazine (C ₆), oxazine (C ₆);

N ₁S ₁: thiazoline (C ₅), thiazolidine (C ₅), parathiazan (C ₆);

N ₂O ₁: oxadiazine (C ₆);

O ₁S ₁: oxygen thia cyclopentenes (C ₅) and oxathiane (thioxane) (C ₆); With

N ₁O ₁S ₁: Evil thiazine (C ₆).

The example of (non-aromatics) the monocyclic heterocycles base that replaces comprises the carbohydrate deutero-group from annular form, for example derived from furanose (C ₅) (for example arbinofuranose, furans lyxose, ribofuranose and furyl xylose) and pyranose (C ₆) (mutter sugar and tower sieve pyranose of A Luo pyranose (allopyranose), A Zhuo pyranose, Glucopyranose, mannopyranose, ancient Lip river pyranose, Chinese mugwort Du pyranose, gala piperazine for example.

Spiral shell C _3-7Cycloalkyl or heterocyclic radical: term used herein " spiral shell C _3-7Cycloalkyl or heterocyclic radical " be meant with another ring and encircle the C that shared single atom is connected by two _3-7Cycloalkyl or C _3-7Heterocyclylalkyl.

Oxygen containing spiral shell C _5-7Heterocyclic radical: term used herein " oxygen containing spiral shell C _5-7Heterocyclic radical " spiral shell C that to relate to 1 annular atoms be oxygen _5-7Heterocyclic radical.

C _5-20Aryl: term " C used herein _5-20Aryl " relate to from C _5-20Remove the univalent perssad that hydrogen atom obtains on the aromatic ring atom of aromatic compound, described compound has a ring, perhaps two or more rings (for example, condensed), and have 5-20 annular atoms, wherein at least one described ring is an aromatic nucleus.Preferably, each ring has 5-7 annular atoms.

Annular atoms can all be a carbon atom, and as in the situation of " carbon aryl ", this group can be called " C easily _5-20Carbon aryl ".

The C that does not have ring hetero atom _5-20Aryl (is C _5-20The carbon aryl) example includes but not limited to from benzene (being phenyl) (C ₆), naphthalene (C ₁₀), anthracene (C ₁₄), luxuriant and rich with fragrance (C ₁₄) and pyrene (C ₁₆) the deutero-group.

Scheme as an alternative, annular atoms can comprise one or more heteroatomss, includes but not limited to oxygen, nitrogen and sulphur, for example in " heteroaryl ".In this case, this group can be called " C easily _5-20Heteroaryl ", " C wherein _5-20" be meant annular atoms, no matter be carbon atom or heteroatoms.Preferably, each ring has 5-7 annular atoms, and wherein 0-4 is ring hetero atom.

C _5-20The example of heteroaryl includes but not limited to from following compounds deutero-C ₅Heteroaryl: furans (oxole), thiophene (dithiole), pyrroles's (azoles), imidazoles (1, the 3-diazole), pyrazoles (1,2-diazole), triazole, oxazole, isoxazole, thiazole, isothiazole, oxadiazole, tetrazolium are with oxatriazole; With from following compounds deutero-C ₆Heteroaryl: Yi oxazine, pyridine (azine), pyridazine (1,2-diazine), pyrimidine (1,3-diazines; For example, cytosine(Cyt), thymus pyrimidine, uridylic), pyrazine (1,4-diazines) and triazine.

Heteroaryl can pass through carbon or heterocyclic atom bonding.

The C that contains condensed ring _5-20The example of heteroaryl includes but not limited to the C derived from cumarone, isobenzofuran, thionaphthene, indoles, isoindole ₉Heteroaryl; C derived from quinoline, isoquinoline 99.9, benzodiazine, pyridopyridine ₁₀Heteroaryl; C derived from acridine and xanthene ₁₄Heteroaryl.

No matter abovementioned alkyl, heterocyclic radical and aryl are separately or as another substituent part, himself can choose wantonly by one or more be selected from they self and following other listed substituting groups replace.

Halogen :-F ,-Cl ,-Br and-I.

Hydroxyl :-OH.

Ether :-OR, wherein R is ether substituting group, for example C _1-7Alkyl (is also referred to as C _1-7Alkoxyl group), C _3-20Heterocyclic radical (is also referred to as C _3-20The heterocyclyloxy base) or C _5-20Aryl (is also referred to as C _5-20Aryloxy), preferred C _1-7Alkyl.

Nitro :-NO ₂

Cyano group (nitrile) :-CN.

(=O) R, wherein R is an acyl substituent to acyl group (ketone) :-C, for example H, C _1-7Alkyl (is also referred to as C _1-7Alkyl acyl or C _1-7Alkyloyl), C _3-20Heterocyclic radical (is also referred to as C _3-20The heterocyclic radical acyl group) or C _5-20Aryl (is also referred to as C _5-20Aryl-acyl), preferred C _1-7Alkyl.The example of acyl group includes but not limited to-C (=O) CH ₃(ethanoyl) ,-C (=O) CH ₂CH ₃(propionyl) ,-C (=O) C (CH ₃) ₃(butyryl radicals) and-C (=O) Ph (benzoyl, benzophenone).

Carboxyl (carboxylic acid) :-COOH.

Ester (carboxylate radical, carboxylicesters, oxygen carbonyl) :-C (=O) OR, wherein R is the ester substituting group, for example, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of ester group includes but not limited to-C (=O) OCH ₃,-C (=O) OCH ₂CH ₃,-C (=O) OC (CH ₃) ₃With-C (=O) OPh.

Aminoacyl (formamyl, carbamyl, aminocarboxyl, carboxylic acid amides (carboxamide)) :-C (=O) NR ¹R ², R wherein ¹And R ²Be amino substituting group independently, as defined to amino.The example of aminoacyl includes but not limited to-C (=O) NH ₂,-C (=O) NHCH ₃,-C (=O) N (CH ₃) ₂,-C (=O) NHCH ₂CH ₃With-C (=O) N (CH ₂CH ₃) ₂, and R ¹And R ²The nitrogen-atoms that connects with their forms the aminoacyl of heterocycle structure, for example amido in piperidino carbonyl, morpholinyl carbonyl, thio-morpholinyl carbonyl and the piperazinyl carbonyl.

Amino :-NR ¹R ², R wherein ¹And R ²Be amino substituting group independently, for example hydrogen, C _1-7Alkyl (is also referred to as C _1-7Alkylamino or two-C _1-7Alkyl oxy), C _3-20Heterocyclic radical or C _5-20Aryl, preferred H or C _1-7Alkyl, or in " ring-type " amino situation, R ¹And R ²The nitrogen-atoms that connects with them forms the heterocycle with 4-8 annular atoms.Amino example includes but not limited to-NH ₂,-NHCH ₃,-NHCH (CH ₃) ₂,-N (CH ₃) ₂,-N (CH ₂CH ₃) ₂With-NHPh.The example of cyclic amino includes but not limited to ethylenimine base, azetidinyl, pyrrolidyl, piperidyl, piperazinyl, perhydro diaza

Base, morpholino base and thiomorpholine are for base.Any substituting group that cyclic amino can be defined by this paper on its ring replaces, for example carboxyl, carboxylate radical and aminoacyl.

Amido (acyl amino) :-NR ¹C (=O) R ², R wherein ¹Be amide substituents, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred H or C _1-7Alkyl, H most preferably, R ²Be acyl substituent, C for example _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of acyl group amido includes but not limited to-NHC (=O) CH ₃,-NHC (=O) CH ₂CH ₃With-NHC (=O) Ph.R ¹And R ²Can form ring texture together, for example in succinimido, maleimide amino and phthalyl imino-:

Urea groups :-N (R ¹) CONR ²R ³, R wherein ²And R ³Be amino substituting group independently, as defined to amino, R ¹Be the urea groups substituting group, for example hydrogen, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred hydrogen or C _1-7Alkyl.The example of urea groups includes but not limited to-NHCONH ₂,-NHCONHMe ,-NHCONHEt ,-NHCONMe ₂,-NHCONEt ₂,-NMeCONH ₂,-NMeCONHMe ,-NMeCONHEt ,-NMeCONMe ₂,-NMeCONEt ₂With-NHC (=O) NHPh.

(=O) R, wherein R is acyloxy substituting group, for example C to acyloxy (anti-ester) :-OC _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of acyloxy includes but not limited to-OC (=O) CH ₃(acetoxyl group) ,-OC (=O) CH ₂CH ₃,-OC (=O) C (CH ₃) ₃,-OC (=O) Ph ,-OC (=O) C ₆H ₄F and-OC (=O) CH ₂Ph.

Sulfydryl :-SH.

Thioether (sulfide) :-SR, wherein R is the thioether substituting group, for example, C _1-7Alkyl (is also referred to as C _1-7Alkylthio), C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.C _1-7The example of alkylthio includes but not limited to-SCH ₃With-SCH ₂CH ₃

Sulfoxide (sulfinyl) :-S (=O) R, wherein R is the sulfoxide substituting group, for example, C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfoxide group includes but not limited to-S (=O) CH ₃With-S (=O) CH ₂CH ₃

Alkylsulfonyl (sulfone) :-S (=O) ₂R, wherein R is sulfone substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfuryl includes but not limited to-S (=O) ₂CH ₃(methyl sulphonyl, methylsulfonyl) ,-S (=O) ₂CF ₃,-S (=O) ₂CH ₂CH ₃With 4-Methyl benzenesulfonyl base (tosyl group).

Sulfo-aminoacyl (thiocarbamyl) :-C (=S) NR ¹R ², R wherein ¹And R ²Be amino substituting group independently, as defined to amino.The example of aminoacyl includes but not limited to-C (=S) NH ₂,-C (=S) NHCH ₃,-C (=S) N (CH ₃) ₂With-C (=S) NHCH ₂CH ₃

Sulfonamido :-NR ¹S (=O) ₂R, wherein R ¹Be amino substituting group, as defined to amino, R is sulfonamido substituting group, for example C _1-7Alkyl, C _3-20Heterocyclic radical or C _5-20Aryl, preferred C _1-7Alkyl.The example of sulfonamido includes but not limited to-NHS (=O) ₂CH ₃,-NHS (=O) ₂Ph and-N (CH ₃) S (=O) ₂C ₆H ₅

As mentioned above, form above-mentioned listed substituent group, for example C _1-7Alkyl, C _3-20Heterocyclic radical and C _5-20Aryl, itself can be substituted.Therefore, substituted substituting group is contained in above-mentioned definition.

Further embodiment

As long as be suitable for, following embodiment can be used for each aspect of the present invention.

In some embodiments, R ^CFor H and Y are CH; In these embodiments, described compound has formula (Ia).

At R ^CBe C _1-4In the embodiment of alkyl, it can be methyl.

If R ^CAnd R ¹Form oxygen containing spiral shell C with carbon atom and Sauerstoffatom that they connected _5-7Heterocyclic radical, then it can be tetrahydrofuran (THF) (it can condense with phenyl ring and form 1,3-dihydro-isobenzofuran).

In some embodiments, Y is CH, and therefore X can be selected from CH, CF and N.

In other embodiments, Y is CF, and therefore X is CH.

In the present invention, the fused aromatic rings that-A-B-represents can only be made up of carboatomic ring atom, therefore can be benzene, naphthalene, and especially can be benzene.As mentioned above, these rings can replace, but are unsubstituted in some embodiments.

In alternate embodiment, the fused aromatic rings that described-A-B-represents can contain the azo-cycle atom, therefore can be, for example, the pyrroles.Therefore compound of the present invention can have formula:

R wherein ^A1, R ^A2And R ^A3Can be independently selected from H and C _1-4Alkyl (for example methyl).In some embodiments, R ^A1And R ^A3In at least one be C _1-4Alkyl (for example methyl).In other embodiments:

(a) R ^A1And R ^A3Be methyl and R ^A2Be hydrogen;

(b) R ^A1, R ^A2And R ^A3Be methyl;

(c) R ^A1Be methyl and R ^A2And R ^A3Be hydrogen.

If the fused aromatic rings that described-A-B-represents has one or more substituting groups, then it can have 2 substituting groups or a divalent substituent.Described substituting group can be connected with the atom on itself being connected center ring α-is connected with carbon atom on the center ring.Therefore, if described fused aromatic rings is a phenyl ring, then the position of substitution is represented with * in following formula in some embodiments:

These substituting groups can be halogen group, especially F.In these embodiments, X can be CH.Described halogen group can also be a chlorine.

In other embodiments, described phenyl ring can be by one or more, and for example two, NH ₂Group replaces.These replacements can occur on the position that following formula points out, and can have one or two substituting group.In some embodiments, described NH ₂On the nearest position of benzyl.

In other embodiments, described phenyl ring can be by one or more, and for example two, C _1-4Alkoxyl group (for example methoxyl group) replaces.These replacements can occur on the position that following formula points out, and can have one or two substituting group.In some embodiments, described alkoxyl group is on the nearest position of benzyl.

In other embodiments, described phenyl ring can be in following formula ⁺Replace on one or two of the position of expression:

These substituting groups can be halogen group, especially chloro or bromo, or NH ₂In some embodiments, there is a substituting group.In other embodiments, there are two these substituting groups.

In some embodiments, X can be CH or CF.Especially, X can be CF.

In other embodiments, X can be N.

Work as R ¹Be C _1-7During alkyl, it can be saturated C _1-7Alkyl, for example methyl, ethyl, sec.-propyl, cyclopropyl methyl.Other examples comprise propyl group, butyl, cyclobutyl and cyclopentyl.It can also be undersaturated, for example propenyl.If described C _1-7Alkyl is substituted, and then substituting group can be selected from above listed substituting group, perhaps can more specifically be: C _5-7Aryl (for example furyl, benzyl, pyridyl), C _3-7Heterocyclic radical (for example tetrahydrofuran base, pyrryl, morpholino base, thiomorpholine are for base), halogen, hydroxyl, C _1-7Alkoxyl group and NH ₂Other substituting groups comprise C _1-4(wherein amino substituting group can be a methyl for alkoxyl group (for example methoxyl group), carboxyl and aminoacyl; perhaps amino substituting group forms heterocyclic radical with the nitrogen-atoms that they connected; morpholino base, 3 for example, 3-two fluoro-azetidinyls, pyrrolidyl or piperidyl).In some embodiments, the C of described replacement _1-7Alkyl is methyl or ethyl.

Work as R ¹Be C _5-20During aryl, it can be C _5-7Aryl.Described aryl can be C ₆Aromatic yl group, for example phenyl or pyridyl.The C that other are possible ₆Aromatic yl group comprises pyridazinyl, pyrimidyl and pyrazinyl.Described aromatic yl group can be unsubstituted or substituted.If described aromatic yl group is substituted, then substituting group can be selected from above listed substituting group, perhaps can more specifically be: C _1-4Alkyl (for example methyl), C _5-7Aryl (for example furyl, benzyl, pyridyl), C _3-7Heterocyclic radical (for example tetrahydrofuran base, pyrryl, morpholino base, thiomorpholine are for base), halogen, hydroxyl, C _1-7Alkoxyl group and NH ₂Other possible substituting groups comprise cyano group.Described substituting group can be selected from halogen (for example F, Cl), hydroxyl and NH ₂, halogen (for example F, Cl) especially.In other embodiments, described substituting group can be selected from halogen, C _1-4Alkoxyl group (for example methoxyl group), cyano group and C _1-4Alkyl (for example methyl).

Work as R ¹Be C _3-20During heterocyclic radical, it can be C _5-7Heterocyclic radical.Described heterocyclic radical group can be, for example, and pyrryl, piperidyl, oxazolyl, isoxazolyl, piperazinyl, morpholinyl and thio-morpholinyl.Described heterocyclic radical group can be unsubstituted or substituted.If described heterocyclic radical is substituted, then substituting group can be selected from above listed substituting group, perhaps can more specifically be: C _5-7Aryl (for example furyl, benzyl, pyridyl), C _3-7Heterocyclic radical (for example tetrahydrofuran base, pyrryl, morpholino base, thiomorpholine are for base), halogen, hydroxyl, C _1-7Alkoxyl group and NH ₂

In some embodiments of the present invention, R ¹Can be methyl or ethyl.

In some embodiments of the present invention, R ¹Can be methyl or ethyl, and R ^CBe H.

In some embodiments of the present invention, R ¹Can be methyl or ethyl, Y be CH, and R ^CBe H.

Other aspects of the present invention are compounds of following examples.

As long as be suitable for, above optimal way is applied in any combination mutually.

Comprise other form

Above comprise these substituent ions of knowing, salt, solvate and protected form.For example, mentioning that carboxylic acid (also comprises its negatively charged ion (carboxylate radical) form (COO COOH) time ^-), salt or solvate and conventional protected form.Similarly, mention the amino amino protonated form (N that also comprises ⁺HR ¹R ²), salt or solvate (for example hydrochloride) and amino conventional protected form.Similarly, mention that hydroxyl also comprises its anionic form (O ^-), the conventional protected form of salt or solvate and hydroxyl.

Isomer, salt, solvate, protected form and prodrug

Some compound can exist with one or more specific geometrical isomers, optical isomer, enantiomer, diastereomer, epimer, steric isomer, tautomer, conformer or anomer form, include but not limited to cis (cis)-with trans (trans)-type; E-and Z-type; C-, t-and r-type; In-with outward-type; R-, S-and meso-type; D-and L-type; D-and l-type; (+) and (-) type; Ketone-, enol-with enolate-type; Along (syn)-with anti-(anti)-type; Synclinal (synclinal)-with anticlinal (anticlinal)-type; α-with β-type; Uprightly (axial) and calm type; Boat form-, chair form-, the distortion formula-, envelope type-with half-chair-type; And combination, below be referred to as " isomer " (or " heterogeneous ").

If compound is a crystallized form, then it can exist with multiple different polymorphic.

Notice that except the tautomeric form of hereinafter discussing, what will get rid of especially is structure (or composition) isomer (that is, the difference of the connection between the atom and be not only the different isomer in atoms in space position) from term used herein " isomer ".For example, mention methoxyl group-OCH ₃Can not be interpreted as mentioning its constitutional isomer methylol-CH ₂OH.Similarly, mention that Chloro-O-Phenyl can not be interpreted as mentioning chloro-phenyl-between its constitutional isomer.But, mention that a class formation then can be included in constitutional isomer form (for example, the C in the type scope _1-7Alkyl comprises n-propyl and sec.-propyl; Butyl just comprising-, different-, secondary-and the tertiary butyl; P-methoxy-phenyl comprise the neighbour-,-and right-p-methoxy-phenyl).

Above-mentioned eliminating does not relate to tautomeric forms, for example ketone, enol and enolate form, for example following tautomer is right: ketone/enol, imines/enamine, acid amides/imino-alcohol, amidine/amidine, nitroso-group/oxime, thioketones/alkene mercaptan, N-nitroso-group/hydroxyl azo-group and nitro/aci-nitro group.

Relevant especially with the present invention is, and tautomer shown below is right:

Note, particularly including term " isomer " in, being compound with one or more isotropic substances replacements.For example, H can be any isotropic substance form, comprises ¹H, ²H (D) and ³H (T); C can be any isotropic substance form, comprises ¹²C, ¹³C and ¹⁴C; O can be any isotropic substance form, comprises ¹⁶O and ¹⁸O etc.

Except as otherwise noted, mention that particular compound comprises the isomeric forms that all are such, comprise (whole or in part) its racemic mixture and other mixtures.The preparation of described isomeric forms (for example asymmetric synthesis) is known in the art with separating (for example fractional crystallization and chromatography) method, perhaps is easy to adopt method teaching herein or currently known methods and obtain according to known way.

Except as otherwise noted, mention that particular compound also comprises its ion and salt form, for example as discussed below.

Except as otherwise noted, mention that particular compound also comprises its solvate forms, for example as discussed below.

Except as otherwise noted, mention that particular compound also comprises its prodrug, for example as discussed below.

Except as otherwise noted, mention that particular compound also comprises its protected form, for example as discussed below.

Except as otherwise noted, mention that particular compound also comprises the polymorphic that they are different, for example as discussed below.

Preparation, purifying and/or handle the corresponding salt of active compound, pharmacologically acceptable salts for example may be easily or desirable.The example of pharmacologically acceptable salts " Berge etc., " Pharmaceutically Acceptable Salts ", J.Pharm.Sci., 66,1-19 discusses in (1977).

For example, if compound is an anionic property, perhaps have can be anionic functional group (for example ,-COOH can be-COO ^-), then can generate salt with the positively charged ion that is fit to.The example of the inorganic cation that is fit to includes but not limited to for example Na of alkalimetal ion ⁺And K ⁺, alkaline earth metal cation is Ca for example ²⁺And Mg ²⁺And other positively charged ions Al for example ³⁺The organic cations example that is fit to includes but not limited to that ammonium ion (is NH ₄ ⁺) and the ammonium ion that replaces (NH for example ₃R ⁺, NH ₂R ₂ ⁺, NHR ₃ ⁺, NR ₄ ⁺).The example of the ammonium ion of the replacement that some is fit to be from following deutero-those: ethamine, diethylamine, dicyclohexyl amine, triethylamine, butylamine, quadrol, thanomin, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine and Trometamol, and amino acid, for example Methionin and arginine.The example of common quaternary ammonium ion is N (CH ₃) ₄ ⁺

If compound is a cationic, perhaps have can be cationic functional group (for example-NH ₂Can be-NH ₃ ⁺), then can generate salt with the negatively charged ion that is fit to.The example of the inorganic anion that is fit to include but not limited to from following mineral acid deutero-those: hydrochloric acid, Hydrogen bromide, hydroiodic acid HI, sulfuric acid, sulfurous acid, nitric acid, nitrous acid, phosphoric acid and phosphorous acid.The example of the organic anion that is fit to include but not limited to from following organic acid deutero-those: acetate, propionic acid, succsinic acid, oxyacetic acid, stearic acid, palmitinic acid, lactic acid, oxysuccinic acid, pounce on acid, tartrate, citric acid, glyconic acid, xitix, toxilic acid, hydroxymaleic acid, toluylic acid, L-glutamic acid, aspartic acid, phenylformic acid, styracin, pyruvic acid, Whitfield's ointment, Sulphanilic Acid, the 2-acetoxy-benzoic acid, fumaric acid, toluenesulphonic acids, methylsulfonic acid, ethyl sulfonic acid, ethionic acid, oxalic acid, isethionic acid, valeric acid and glyconic acid.The anionic example of polymerization that is fit to include but not limited to from following polymers acid deutero-those: tannic acid, carboxymethyl cellulose.

Preparation, purifying and/or the coordinative solvent thing of handling active compound may be easily or desirable.Term used herein " solvate " is meant the mixture of solute (for example salt of active compound, active compound) and solvent on conventional meaning.If solvent is a water, then solvate can be called hydrate, for example monohydrate, dihydrate, trihydrate etc. easily.

Preparation, purifying and/or the active compound of handling the chemoproection form may be easily or desirable.It is protected and avoid the compound of undesirable chemical reaction that term used herein " chemoproection form " relates to one or more reactive functional groups, and promptly compound is protected or the form of blocking group (be also referred to as masked or shelter group or be closed or blocking groups).By protective reaction functional group, can relate to the reaction of other unprotected reactive functional groups, and not influence protected group; Blocking group is removed in step subsequently usually, and does not influence the remainder of molecule substantially.Referring to, for example, " Protective Groups in Organic Synthesis " (T.Green and P.Wuts; 3rd Edition; John Wiley and Sons, 1999).

For example, hydroxyl can protectedly be ether (OR) or ester (OC (=O) R), for example, tertbutyl ether; Benzyl, diphenyl-methyl (diphenyl methyl) or trityl (trityl group) ether; Trimethyl silyl or t-butyldimethylsilyl ether; Or ethanoyl ester (OC (=O) CH ₃,-OAc).

For example, the aldehydes or ketones group can be protected as acetal or ketal respectively, wherein carbonyl (＞C=O) by being converted into diether (＞C (OR) with the reaction of primary alconol for example ₂).Aldehyde radical or ketone group are easy to by using a large amount of excessive water hydrolysis to regenerate in the presence of acid.

For example, amine groups for example can be used as acid amides or urethane is protected, for example methyl nitrosourea (NHCO-CH ₃); Benzyloxy acid amides (NHCO-OCH ₂C ₆H ₅,-NH-Cbz); Tert.-butoxy acid amides (NHCO-OC (CH ₃) ₃,-NH-Boc); 2-biphenyl-2-propoxy-acid amides (NHCO-OC (CH ₃) ₂C ₆H ₄C ₆H ₅,-NH-Bpoc); 9-fluorenyl methoxy acid amides (NH-Fmoc); 6-nitro black false hellebore oxygen base acid amides (NH-Nvoc); 2-trimethylsilylethoxy) acid amides (NH-Teoc); 2,2,2-three chloroethoxy acid amides (NH-Troc); The allyloxy acid amides (NH-Alloc); (2-benzenesulfonyl) the oxyethyl group acid amides (NH-Psec); Perhaps under situation about being fit to, as the N-oxide compound (＞NO) protect.

For example, hydroxy-acid group can be used as ester or acid amides is protected, described ester for example: C _1-7Alkyl ester (for example methyl ester, tertiary butyl ester); C _1-7Haloalkyl ester (C for example _1-7Three alkylhalide group esters); Three C _1-7Alkyl silyl-C _1-7Alkyl ester; C _5-20Aryl-C _1-7Alkyl ester (for example benzyl ester, nitrobenzyl ester); Described acid amides is methyl nitrosourea for example.

For example, sulfydryl can be used as thioether and (SR) protects, for example: the benzyl thioether; Acetylamino methyl ether (S-CH ₂NHC (=O) CH ₃).

Preparation, purifying and/or the active compound of handling prodrug forms may be easily or desirable.Term used herein " prodrug " is meant when carrying out metabolism and generates the compound of desirable active compound when (for example in vivo).Usually, prodrug is a non-activity, and perhaps its activity is lower than active compound, but favourable operation, administration or metabolic characteristic can be provided.

For example, some prodrug is the ester (the unsettled ester of for example physiologically acceptable metabolism) of active compound.In metabolic process, ester group (C (=O) OR) cracking generates active medicine.Described ester can be by for example forming the esterification of any one hydroxy-acid group in the parent compound (C (=O) OH); in appropriate circumstances; can protect any other reactive group that exists in the parent compound earlier, and then deprotection as required.The example of the unsettled ester of this metabolism comprise following these: wherein R is C _1-20Alkyl (for example-Me ,-Et); C _1-7Aminoalkyl group (for example amino-ethyl, 2-(N, N-diethylamino) ethyl, 2-(4-morpholino) ethyl); And acyloxy-C _1-7Alkyl (acyloxy methyl for example, acyloxy ethyl, for example oxy acid methyl neopentyl, acetoxy-methyl, 1-acetoxyl group ethyl, 1-(1-methoxyl group-1-methyl) ethyl-ketonic oxygen base ethyl, 1-(benzoyloxy) ethyl, isopropoxy-ketonic oxygen ylmethyl, 1-isopropoxy-ketonic oxygen base ethyl, cyclohexyl-ketonic oxygen ylmethyl, 1-cyclohexyl-ketonic oxygen base ethyl, cyclohexyl oxygen base-ketonic oxygen ylmethyl, 1-cyclohexyl oxygen base-ketonic oxygen base ethyl, (4-THP trtrahydropyranyl oxygen base) ketonic oxygen ylmethyl, 1-(4-THP trtrahydropyranyl oxygen base) ketonic oxygen base ethyl, (4-THP trtrahydropyranyl) ketonic oxygen ylmethyl and 1-(4-THP trtrahydropyranyl) ketonic oxygen base ethyl) ester.

Other prodrug forms that are fit to comprise phosphonic acid ester and oxyacetate.Particularly, and hydroxyl (OH) can be by reacting with the chloro dibenzyl phosphite, hydrogenation forms phosphonate groups-O-P (=O) (OH) then ₂And be prepared into phosphonate prodrugs.This group can be removed by phosphoesterase in metabolic process and generate the active medicine with hydroxyl.

In addition, some prodrug is generated active compound by the enzyme catalysis activation, or generates the compound of active compound through further chemical reaction.For example, prodrug can be sugar derivatives or other glucosides binding substances, perhaps can be amino acid ester derivative.

Acronym

For simplicity; many chemical groups are represented with the abbreviation of knowing, are included but not limited to methyl (Me), ethyl (Et), n-propyl (nPr), sec.-propyl (iPr), normal-butyl (nBu), the tertiary butyl (tBu), n-hexyl (nHex), cyclohexyl (cHex), phenyl (Ph), xenyl (biPh), benzyl (Bn), naphthyl (naph), methoxyl group (MeO), oxyethyl group (EtO), benzoyl (Bz) and ethanoyl (Ac).

For simplicity, chemical compound lot is represented with the abbreviation of knowing, is included but not limited to methyl alcohol (MeOH), ethanol (EtOH), Virahol (i-PrOH), methylethylketone (MEK), ether or Anaesthetie Ether (Et ₂O), acetate (AcOH), methylene dichloride (METHYLENE CHLORIDE, DCM), trifluoroacetic acid (TFA), dimethyl formamide (DMF), tetrahydrofuran (THF) (THF) and dimethyl sulfoxide (DMSO) (DMSO).

Synthetic

Compound of the present invention can be by making the compound of formula 1:

Wherein A, B and X such as preamble define,

Compound with formula 2:

Wherein R1 such as preamble define,

At coupling agent system 2-(1H-benzotriazole-1-yl)-1 for example, 1,3,3-tetramethyl-urea a tetrafluoro borate, 2-(1H-benzotriazole-1-yl)-1,1,3, under the existence of 3-tetramethyl-urea hexafluorophosphate or (dimethylamino-propyl) second carbonyl inferior amine salt hydrochlorate/hydroxybenzotriazole, alkali for example diisopropylethylamine in the presence of, for example synthesize at 0 ℃ of temperature range internal reaction in N,N-DIMETHYLACETAMIDE or the methylene dichloride at solvent to the solvent for use boiling point.

As alternative scheme, compound of the present invention can adopt the method for knowing to be converted into active substance, for example acyl chlorides, or activatory ester by the compound with formula 1, N-hydroxyl amber imines ester for example, and make the compound reaction of described active substance and formula 2 come synthetic.

It is that CH and X are compound synthetic of the formula 1 of CH or CF that WO 2004/080976 has described Y, and its description is incorporated herein by reference.It is that CH and X are compound synthetic of the formula 1 of N that WO 2006/021801 has described Y, and its description is incorporated herein through quoting.

The compound of formula 2 is commercially available, perhaps can be synthetic by reported method in the chemical literature.

R ^CThe compound that is the formula 2 of H can also be by the compound of formula 3:

R wherein ¹Define as preamble,

Appropriate catalyst for example carrying alumina 5% rhodium in the presence of, for example synthesize by hydrogenation in the ethanol in suitable solvent.

The compound of formula 3 is commercially available, perhaps can be synthetic by reported method in the chemical literature.

The compound of formula 1 can also be by making the compound of formula 4:

Wherein A, B and X such as preamble define, and Q is suitable leavings group, bromine for example,

In the presence of carbon monoxide and appropriate catalyst, react to come synthetic, described catalyzer is for example trans-two [2-(di-o-tolyl phosphine) benzyl] acetate two palladiums (II) of two-mu-.

The compound of formula 4 can synthesize by the method that preamble is quoted in WO 2004/080976 and WO 2006/021801, perhaps by making the compound of formula 5:

Wherein A, B, X, Y and Q such as preamble define,

The perhaps compound of formula 6 (and/or its relevant open loop form formula 7):

Wherein A, B, X, Y and Q such as preamble define,

Perhaps the mixture of formula 5,6 and 7 compound alkali for example triethylamine in the presence of, perhaps acid for example acetate in the presence of, randomly solvent for example water, DMF or THF in the presence of for example hydrazine hydrate or hydrazine hydrate reaction come synthetic with the hydrazine source.

The compound of formula 5 can be by synthesizing with the method similar methods described in WO 2004/080976 and the WO 2006/021801, perhaps by making the compound of formula 8:

Wherein A and B such as preamble define,

Compound with formula 9:

Wherein X, Y and Q such as preamble define,

Alkali for example sodium acetate in the presence of, reaction comes synthetic under the temperature on the fusing point of used compound.

The compound of the compound of formula 8 and formula 9 is commercially available, perhaps can synthesize by the chemical literature reported method.

In another embodiment, compound of the present invention can itself be a compound of the present invention by making the compound of formula 10:

Wherein A, B, X and Y such as preamble define,

Compound with formula 11:

T-R ¹Formula 11

R wherein ¹Define as preamble, typically be the optional C that replaces ₆-heteroaryl, T are leavings groups, fluorine or chlorine for example,

Suitable alkali for example sodium hydride or sodium tert-butoxide in the presence of react synthetic.

In addition, compound of the present invention can make the compound of formula 10 by the arylation reaction, and wherein A, B, X and Y such as preamble define, with the compound of formula 11, wherein R ¹Define as preamble, typically be the optional C that replaces ₆-heteroaryl, and Y is hydroxyl reacts synthetic, and described arylation reaction relates to uses triphenylphosphine and diethylazodicarboxylate.

In other embodiments, the compound that compound of the present invention can be by making formula 12 or the compound of formula 13:

Wherein A, B, X and Y such as preamble define, and it is compound of the present invention, and carboxylic acid functional can be randomly esterified, for example with the form of methyl esters,

With the hydrazine source, for example a hydrazine hydrate or a hydrazine hydrate, randomly alkali for example triethylamine or acid for example acetate in the presence of, randomly solvent for example water, DMF or THF in the presence of react synthetic.

The compound of formula 12 can be by making the compound of formula 8:

Wherein A and B such as preamble define,

Compound with formula 14:

Wherein X, Y and R ¹Define as preamble,

Alkali for example sodium acetate in the presence of, under the temperature on the fusing point of compound used therefor, react to come synthetic.

The compound of formula 14 can be by the compound of formula 15:

Wherein X, Y and R ¹Define as preamble, and K is a leavings group, typically is iodo,

By appropriate catalyst for example cuprous iodide (I) and suitable alkali for example cesium carbonate in the presence of, come syntheticly with the diethyl malonate reaction, for example in suitable solvent such as THF/ water, use alkali such as lithium hydroxide to carry out decarboxylation and ester hydrolysis then.

The compound of formula 15 can be by making the compound of formula 16:

Wherein X, Y and Q such as preamble define,

Compound with formula 2:

R wherein ¹Define as preamble,

At coupling agent system 2-(1H-benzotriazole-1-yl)-1 for example, 1,3,3-tetramethyl-urea a tetrafluoro borate, 2-(1H-benzotriazole-1-yl)-1,1,3, under the existence of 3-tetramethyl-urea hexafluorophosphate or (dimethylamino-propyl) second carbonyl inferior amine salt hydrochlorate/hydroxybenzotriazole, alkali for example diisopropylethylamine in the presence of, for example synthesize at 0 ℃ of temperature range internal reaction in N,N-DIMETHYLACETAMIDE or the methylene dichloride in solvent to the solvent for use boiling point.

The compound of the compound of formula 8 and formula 16 is commercially available, perhaps can be synthetic by reported method in the chemical literature.

The compound of formula 13, especially A and Type B:

Wherein X, Y and R ¹Define as preamble,

Can be by the compound of formula 17:

Compound with formula 14

Wherein X, Y and R ¹Define as preamble,

Carry out acylation reaction and synthesize, earlier the compound with formula 14 is converted into active substance, chloride of acid for example, then Lewis acid for example aluminum chloride in the presence of react.

The compound of formula 17 is commercially available, perhaps can be synthetic by reported method in the chemical literature.

Purposes

The invention provides active compound, specifically is to suppress the active active compound of PARP.

Term used herein " activity " relates to can suppress the active compound of PARP, specifically comprises the prodrug of compound (medicine) with intrinsic activity and such compound, and described prodrug itself can show seldom or not show intrinsic activity.

In order to estimate the PARP restraining effect that particular compound provides, hereinafter embodiment has described a kind of assay method that can conveniently use.

The present invention also provides and suppresses the active method of PARP in the cell, comprises described cell is contacted with the active compound of significant quantity, and described active compound preferably pharmacy can be accepted the form of composition.This method can be implemented in external or body.

For example, can make cell sample, observe the effect of compound these cells in growth in vitro and with active compound and described cells contacting.As the example of " effect ", can measure the amount that the interior sometime DNA of realization repairs.When finding that the active compound pair cell exerts an influence, it in carrying patient's the method for cell of same cell type, can be used as in treatment the prognostic markers thing or the diagnostic marker of compound efficacy.

With regard to the treatment illness, term used herein " treatment " is usually directed to obtain for the mankind or animal (for example in the animal doctor uses) treatment or the therapy of some desired therapeutic effect, described desired therapeutic effect for example suppresses illness progress, comprises the stopping of decline, progression rates of progression rates, the improvement of illness and the healing of illness.Also comprise treatment (i.e. prevention) as preventive measures.

Term used herein " auxiliary " is meant active compound and known treatment means combined utilization.These means comprise cytotoxic drug therapy and/or the ionizing rays when being used for the treatment of the various cancers type.Particularly, the known activity compound can strengthen the effect of multiple cancer chemotherapy therapeutical agent, and described therapeutical agent comprises the drug toxicity (for example topotecan, Rinotecan and rubitecan) of the topoisomerase enzyme that is used for the treatment of cancer, most of known alkylating agent (for example DTIC, Temozolomide) and based on the medicine (for example carboplatin, cis-platinum) of platinum.

Active compound also can be as the cell cultures additive to suppress PARP, for example in order to make cell external responsive more to known chemotherapeutic or ionizing radiation treatment agent.

Active compound also can be as the part of external test method, for example in order to determine whether candidate host may be benefited from the compounds for treating of being discussed.

Administration

Active compound or the pharmaceutical composition that comprises active compound can be administered to the experimenter by any suitable route of administration, and no matter general/periphery includes but not limited to still at the site of action of expectation: oral (for example by taking in); Local (for example comprise in transdermal, the nose, intraocular, oral cavity and hypogloeeis); Lung (for example, use aerosol for example by for example mouthful the suction of nose or be blown into therapy); Rectum; Vagina; Parenteral, for example by injection, comprise in subcutaneous, intracutaneous, intramuscular, intravenously, intra-arterial, intracardiac, the sheath, in the backbone, in the capsule, under the capsule, in the eye socket, in the intraperitoneal, tracheae, under the epidermis, intraarticular, arachnoid membrane be down and in the breastbone; By implanting Drug Storage, for example subcutaneous or intramuscular.

The experimenter can be eukaryote, animal, vertebrates, Mammals, rodent (for example cavy, hamster, rat, mouse), murine (for example home mouse), Canis animals (for example dog), feline (for example cat), equine species (for example horse), primates, man like ape (for example monkey or ape), monkey class (for example marmoset, baboon), apes (for example gorilla, chimpanzee, orangutan, gibbon) or the mankind.

Preparation

Although can use active compound separately, preferably it is as containing aforesaid at least a active compound and one or more pharmaceutically acceptable carriers, adjuvant, vehicle, thinner, weighting agent, buffer reagent, stablizer, sanitas, lubricant or other materials well known to those skilled in the art and optional other treatment agent or the pharmaceutical composition (as preparation) of preventive.

Therefore, the present invention also provides the method for pharmaceutical composition and pharmaceutical compositions as defined above, and described method comprises mixes at least a active compound as defined above with one or more pharmaceutically acceptable carriers, vehicle, buffer reagent, adjuvant, stablizer or other materials described herein.

Term used herein " pharmacy is acceptable " relates to and is suitable for contacting with experimenter's (for example human) tissue in the scope that rational medicine is judged and does not have compound, material, composition and/or a formulation of excessive toxicity, pungency, anaphylaxis or other problems or complication, its be equivalent to rational benefited/the risk ratio.Every kind of carrier, vehicle etc. must also be " acceptable " on the meaning compatible with other compositions of preparation.

Suitable carriers, thinner, vehicle etc. can find in the standard pharmaceutical textbook.Referring to, for example " Handbook of Pharmaceutical Additives ", the 2nd edition (M.Ash and I.Ash compile), 2001 (Synapse Information Resources, Inc., Endicott, New York, USA), " Remington ' s Pharmaceutical Sciences ", the 20th edition, Lippincott, Williams ﹠amp; Wilkins publishes, and 2000; " Handbook of Pharmaceutical Excipients ", the 2nd edition, 1994.

Preparation can adopt the form of unit dosage form easily, and can prepare by any method that pharmaceutical field is known.These methods comprise the step of active compound with the carrier combinations that constitutes one or more ancillary components.Generally speaking, described preparation by with active compound with liquid vehicle or solid carrier in small, broken bits or with both evenly, mix fully, formed product is prepared.

Preparation can be liquid, solution, suspensoid, emulsion, elixir, syrup, tablet, lozenge, granule, powder, capsule, cachet, pill, injection, suppository, vaginal suppository, ointment, gelifying agent, paste, ointment, sprays, aerosol, foaming agent, lotion, finish, bolus, electuary or aerosol.

The preparation that is suitable for oral administration (for example by taking in) can be a discrete unit, for example: capsule, cachet or tablet, each all contains the active compound of predetermined amount; Powder or granule; Solution in waterborne liquid or non-aqueous liquid or suspensoid; Or oil-in-water liq emulsion or water-in-oil-type liquid emulsion; Bolus; Electuary; Or paste.

Tablet can pass through ordinary method, for example randomly with one or more ancillary component compressing tablets or molded the preparation.The tablet of compacting can by in suitable machine with the free-flowing form active compound of powder or particle form for example, optional and one or more tackiness agents are (as polyvidone, gelatin, gum arabic, Sorbitol Powder, tragacanth gum, Vltra tears), weighting agent or thinner are (as lactose, Microcrystalline Cellulose, secondary calcium phosphate), lubricant is (as Magnesium Stearate, talcum powder, silicon-dioxide), disintegrating agent is (as primojel, polyvinylpolypyrrolidone, croscarmellose sodium), tensio-active agent or dispersion agent or wetting agent (as sodium lauryl sulphate) and sanitas are (as methyl p-hydroxybenzoate, propylparaben, Sorbic Acid) mixing prepares.Molded tablet can prepare by molded mixture with the moistening powder compound of inert liquid diluent in suitable machine.Tablet can randomly carry out dressing or impression, and can prepare tablet with the releasing properties that expectation is provided by for example using the Vltra tears of different ratios, with slowly-releasing or the controlled release that active compound is provided.Tablet can be chosen wantonly has enteric coating to be provided at enteron aisle part rather than in the release of stomach.

The preparation (for example in the transdermal, nose, intraocular, oral cavity and hypogloeeis) that is suitable for topical can be prepared as ointment, ointment, suspensoid, lotion, powder, solution, paste, gelifying agent, sprays, aerosol or finish.Scheme as an alternative, preparation can contain paster or dressing and for example be soaked with active compound and choose any one kind of them or the bandage or the viscosity plaster of multiple vehicle or thinner.

The preparation that is suitable for mouthful interior topical comprises: lozenge comprises active compound in its matrix in seasoning (being generally sucrose and gum arabic or tragacanth gum); Pastille, it comprises active compound in inert base (for example gelatin and glycerine or sucrose and gum arabic); And mouth wash shua, it comprises active compound in the liquid vehicle that is fit to.

The preparation that is suitable for the eye topical also comprises eye drop, and wherein active compound is dissolved in or is suspended in the suitable carriers, in the aqueous solvent especially for active compound.

Carrier is that solid is applicable to that the preparation of nose administration comprises that particle size is the about 500 microns coarse meal of for example about 20-, and it smells the mode administration of agent with employing, and the container that is about to be equipped with powder sucks by nasal meatus fast near nose.Carrier is being used for for example nasal spray, nasal drop administration or comprising the water-based or the oily solution agent of active compound by atomizer with the appropriate formulation of aerosol drug delivery of liquid.

The preparation that is adapted to pass through inhalation comprises the preparation that produces sprays from pressurized package, uses suitable aerosol propellant such as Refrigerant 12, trichlorofluoromethane, dichloro tetrafluoro ethane, carbonic acid gas or other suitable gas.

Be suitable for comprising ointment, ointment and emulsion through the preparation of local skin administration.When being formulated as ointment, active compound is optional to be used with paraffin or with the miscible ointment base of water.Scheme as an alternative, active compound can be mixed with ointment with the oil-in-water-type emulsifiable paste matrix.If necessary, the water of emulsifiable paste matrix for example can comprise at least about the polyvalent alcohol of 30%w/w, promptly has the alcohol of two or more hydroxyls such as propylene glycol, fourth-1,3-glycol, N.F,USP MANNITOL, sorbyl alcohol, glycerine and polyoxyethylene glycol and composition thereof.Topical formulations can desirably comprise the enhanced activity compound by skin or the absorption of other affected area or the compound of infiltration.The example of such dermal osmosis reinforcer comprises dimethyl sulfoxide (DMSO) and related analogs.

When being prepared as local emulsion, oil phase can be chosen wantonly and only comprise emulsifying agent (being called emulgent in addition), and perhaps it can comprise at least a emulsifying agent and oily or the fatty and oily mixture of fat.Preferably, comprise hydrophilic emulsifier and with the lipophilic emulsifier of used as stabilizers.Also preferred package oil-containing and fat.In general, the emulsifying agent that contains stablizer or do not contain stablizer is formed so-called emulsifying wax, and this wax is formed so-called emulsification ointment base with oil and/or fat, thereby constitutes the oily disperse phase of ointment.

Suitable emulgent and emulsion stablizer comprise tween (Tween) 60, sapn (Span) 80, cetostearyl alcohol, tetradecyl alcohol, glyceryl monostearate and Sodium Lauryl Sulphate BP/USP.Be suitable for the oil of preparation or fatty selection based on realizing required beauty treatment character, because the solubleness of active compound in most of oil that may be used in pharmaceutical emulsion may be very low.Therefore preferred greasy, the product that do not dye and can wash off of right and wrong of ointment, its have suitable viscosity with avoid from manage or other containers seepage.Can use straight or branched monobasic or binary alkyl ester for example two dissidents, two acid esters, iso-spermaceti ester alcohol stearic acid, coconut fatty acid propylene glycol diesters, Isopropyl myristate, decyl oleate, Wickenol 111, butyl stearate, palmitinic acid 2-(ethyl hexyl) ester or be called the mixture of the branched ester of Crodamol CAP, last three is preferred ester.These can use separately or use according to required properties of combination.Perhaps, can use high-melting-point lipid such as paraffinum molle alba and/or whiteruss or other mineral oil.

The preparation that is applicable to rectal administration can be the form with suppository of suitable matrix, and suitable matrix contains for example theobroma oil or salicylate.

The preparation that is applicable to vagina administration can be to contain the form that also contains vaginal suppository, cotton balls, emulsifiable paste, gel, paste, foaming agent or the sprays of suitable carrier known in the art except that the active ingredient beyond the region of objective existence.

The preparation that is applicable to parenteral admin is (for example by injection, comprise intracutaneous, subcutaneous, intramuscular, intravenously and intradermal) comprising: water-based and nonaqueous etc. is oozed, pyrogen-free aseptic injectable solution agent, and it can contain antioxidant, buffer reagent, sanitas, stablizer, fungistat and make preparation and the isoosmotic solute of experimenter's blood; Water-based and nonaqueous aseptic suspensoid, it can comprise suspending agent and thickening material, and liposome or be designed for other microparticulate systems that make targeting compounds blood constitutent or one or more organs.The example that the suitable grade that is used for this preparation is oozed carrier comprises sodium chloride injection, Ringer's solution, ringer's lactate injection liquid.Usually, the concentration of active compound in solution is the about 10 μ g/ml of about 1ng/ml-, the about 1 μ g/ml of for example about 10ng/ml-.Preparation can be the form of unitary dose or multiple doses sealed vessel, for example ampoule and phial, and can be stored under lyophilize (freeze-drying) condition, only need add at once before use sterile liquid carrier for example water for injection get final product.Can be from sterilized powder, particle and tablet preparation interim injection solution and suspension.Preparation can be that liposome or design make the form of active compound target in other microparticulate systems of blood constitutent or one or more organs.

Dosage

Should be appreciated that active compound and the suitable dose that contains the composition of active compound can be different because of the patient.Determine that optimal dose is usually directed to weigh between any risk for the treatment of interests level and the present invention treatment or harmful side effect.Selected dosage level will depend on various factors, include but not limited to activity, route of administration, administration time, the discharge rate of compound, the time length of treatment, the other drug that is used in combination, compound and/or material and patient's age, sex, body weight, illness, general health situation and the medical history of particular compound.The amount of compound and route of administration are finally by doctor's decision, but common described dosage should be able to obtain the partial concn at action site place with the realization desired effects, and do not cause the harmful or toxic side effect of essence.

Be applied in the body in the whole course of treatment can with single dose, continuously or intermittently (for example at the dosage of proper spacing) to separate carry out.The method of the most effective administering mode and dosage of determining is known for those skilled in the art, and can change along with the preparation that is used for the treatment of, therapeutic purpose, the target cell of being treated, the experimenter that treated.Can carry out the single or multiple administration, dosage level and pattern are selected by the treatment doctor.

Generally speaking, the suitable dose of active compound is the about 250mg of every kg experimenter's body weight about 100 μ g-every day.If active compound is salt, ester, prodrug etc., the amount of using is basic calculation with the parent compound, and therefore used actual weight can increase pro rata.

Polymorphic

Following crystal formation is hereinafter to prepare among the embodiment 47.

The compound 2b of crystal form A (anhydrous)

Compound 2b (anhydrous crystal forms A) is characterised in that to have the following 2 θ values that at least one uses the CuK alpha-ray to record: 19.9 ° and 4.9 °.Compound 2b (anhydrous crystal forms A) is characterised in that the X-ray powder diffraction pattern that has basically as shown in Figure 1.Listed 10 main peaks in the table 1:

Table 1

10 X-ray powder diffraction main peaks of the anhydrous compound 2b of crystal form A

2-θ angle (2 θ)	Intensity %	Relative intensity
			4.9	60	vs
9.9	17	s
			13.2	13	s
14.9	15	s
			15.5	19	s
17.4	40	vs
			17.8	13	s
19.9	100	vs
			24.4	12	s
24.9	10	s

Vs=is extremely strong

S=is strong

Therefore, according to another aspect of the present invention, provide a kind of compound 2b of crystal formation, anhydrous crystal forms A, its X-ray powder diffraction pattern have at least one characteristic peak about 2-θ=19.9 °.

According to a further aspect in the invention, provide a kind of compound 2b of crystal formation, anhydrous crystal forms A, its X-ray powder diffraction pattern have at least one characteristic peak about 2-θ=4.9 °.

According to a further aspect in the invention, provide a kind of compound 2b of crystal formation, anhydrous crystal forms A, its X-ray powder diffraction pattern have at least two characteristic peaks about 2-θ=19.9 ° and 4.9 °.

According to a further aspect in the invention, a kind of compound 2b of crystal formation is provided, anhydrous crystal forms A, its X-ray powder diffraction pattern have the characteristic peak about 2-θ=4.9 °, 9.9 °, 13.2 °, 14.9 °, 15.5 °, 17.4 °, 17.8 °, 19.9 °, 24.4 ° and 24.9 °.

According to a further aspect in the invention, provide a kind of compound 2b of crystal formation, anhydrous crystal forms A, its X-ray powder diffraction pattern and X-ray powder diffraction pattern shown in Figure 1 are basic identical.

Dsc analysis shows that the compound 2b of anhydrous crystal forms A is a high melting solid, and it and melting peak (Fig. 2) occurs at 143 ℃ 134 ℃ of beginning fusions when with 10 ℃/minute speed heating.

When mentioning that aspect of the present invention relates to the crystal formation of compound 2b, degree of crystallinity is suitably greater than about 60%, more suitably greater than about 80%, be preferably greater than about 90%, more preferably greater than about 95%.Most preferably degree of crystallinity is greater than about 98%.

The compound 2b of anhydrous crystal forms A has and the essentially identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 1, and has 10 main peaks shown in the table 1 (2-θ angle value) substantially.Should be appreciated that the 2-θ angle value for different instruments or different sample X-ray powder diffraction pattern can change slightly, thus aforementioned value can not to be interpreted as be absolute.

As everyone knows, the X-ray powder diffraction pattern that (for example instrument of Shi Yonging or equipment) can obtain having one or more measuring error under different measuring conditions.Particularly, known usually, the intensity of X-ray powder diffraction pattern may fluctuate under different measuring conditions.Therefore, be to be understood that, the 2b of crystal form A of the present invention is not limited to have the crystal with the identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 1, and any have with the crystal of the essentially identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 1 all within the scope of the invention.The technician in X-ray powder diffraction field can judge the basically identical of X-ray powder diffraction pattern.

Crystal form B (hydrate) compound 2b

Compound 2b (hydrate crystal forms B) is characterised in that to have at least one that use following 2 θ values that the CuK alpha-ray records: 21.7 ° and 16.5 °.2b (hydrate crystal forms B) is characterised in that the X-ray powder diffraction pattern that has basically as shown in Figure 3.Listed 10 main peaks in the table 2:

Table 2

10 X-ray powder diffraction main peaks of the compound 2b of hydrate crystal forms B

2-θ angle (2 θ)	Intensity %	Relative intensity
			9.2	27	vs
11.9	68	vs
			16.5	82	vs
18.2	29	vs
			19.7	47	vs
21.4	46	vs
			21.7	100	vs

22.1	23	s
			23.9	42	vs
28.4	25	vs

Vs=is extremely strong

S=is strong

Therefore, according to a further aspect in the invention, provide the crystal formation of a kind of compound 2b, hydrate crystal forms B, its X-ray powder diffraction pattern have at least one characteristic peak about 2-θ=21.7 °.

According to a further aspect in the invention, provide the crystal formation of a kind of compound 2b, hydrate crystal forms B, its X-ray powder diffraction pattern have at least one characteristic peak about 2-θ=16.5 °.

According to a further aspect in the invention, provide the crystal formation of a kind of compound 2b, hydrate crystal forms B, its X-ray powder diffraction pattern have at least two characteristic peaks about 2-θ=21.7 ° and 16.5 °.

According to a further aspect in the invention, the crystal formation of a kind of compound 2b is provided, hydrate crystal forms B, its X-ray powder diffraction pattern have the characteristic peak about 2-θ=9.2 °, 11.9 °, 16.5 °, 18.2 °, 19.7 °, 21.4 °, 21.7 °, 22.1 °, 23.9 ° and 28.4 °.

According to a further aspect in the invention, provide the crystal formation of a kind of compound 2b, the hydration crystal form B, its X-ray powder diffraction pattern and X-ray powder diffraction pattern shown in Figure 3 are basic identical.

Dsc analysis shows that the compound 2b of hydrate crystal forms B is a solid, and it begins the heat absorption dehydration at 31 ℃ when with 10 ℃/minute speed heating, and at 105 ℃ of beginning second endothermic transitions (Fig. 4).

When mentioning that aspect of the present invention relates to a kind of compound 2b of crystal formation, promptly during hydrate crystal forms B, degree of crystallinity is suitably greater than about 60%, more suitably greater than about 80%, be preferably greater than about 90%, more preferably greater than about 95%.Most preferably degree of crystallinity is greater than about 98%.

The compound 2b of hydrate crystal forms B has and the essentially identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 3, and has 10 main peaks shown in the table 2 (2-θ angle value) substantially.Should be appreciated that the 2-θ angle value for different instruments or different sample X-ray powder diffraction pattern can change slightly, thus aforementioned value can not to be interpreted as be absolute.

As everyone knows, the X-ray powder diffraction pattern that (for example instrument of Shi Yonging or equipment) can obtain having one or more measuring error under different measuring conditions.Particularly, known usually, the intensity of X-ray powder diffraction pattern may fluctuate under different measuring conditions.Therefore, be to be understood that, the compound 2b of hydrate crystal forms B of the present invention is not limited to have the crystal with the identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 3, and any have with the crystal of the essentially identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 3 all within the scope of the invention.The technician in X-ray powder diffraction field can judge the basically identical of X-ray powder diffraction pattern.

The compound 2f of crystal form A (anhydrous)

Compound 2f (anhydrous crystal forms A) is characterised in that to have at least one that use following 2 θ values that the CuK alpha-ray records: 19.9 ° and 4.9 °.Compound 2f (crystal form A) is characterised in that the X-ray powder diffraction pattern that has basically as shown in Figure 5.Listed 10 main peaks in the table 3:

Table 3

10 X-ray powder diffraction main peaks of the compound 2f of crystal form A

2-θ angle (2 θ)	Intensity %	Relative intensity
			19.9	100.0	vs
4.9	64.5	vs
			17.4	41.8	vs
15.5	20.4	s
			9.9	17.5	s
14.9	15.3	s
			13.2	14.2	s
17.8	13.8	s
			24.4	12.7	s
24.9	10.5	s

Vs=is extremely strong

S=is strong

Among the m=

A little less than the w=

Therefore, according to a further aspect in the invention, provide a kind of compound 2f of crystal formation, crystal form A, its X-ray powder diffraction pattern have at least one characteristic peak about 2-θ=19.9 °.

According to a further aspect in the invention, provide a kind of compound 2f of crystal formation, crystal form A, its X-ray powder diffraction pattern have at least one characteristic peak about 2-θ=4.9 °.

According to a further aspect in the invention, provide a kind of compound 2f of crystal formation, crystal form A, its X-ray powder diffraction pattern have at least two characteristic peaks about 2-θ=19.9 ° and 4.9 °.

According to a further aspect in the invention, a kind of compound 2f of crystal formation is provided, crystal form A, its X-ray powder diffraction pattern have the characteristic peak about 2-θ=4.9 °, 9.9 °, 13.2 °, 14.9 °, 15.5 °, 17.4 °, 17.8 °, 19.9 °, 24.4 ° and 24.9 °.

According to a further aspect in the invention, provide a kind of compound 2f of crystal formation, crystal form A, its X-ray powder diffraction pattern and X-ray powder diffraction pattern shown in Figure 5 are basic identical.

Dsc analysis shows that the compound 2f of crystal form A is a high melting solid, and it is 116 ℃ of beginning fusions (Fig. 6) when with 10 ℃/minute speed heating.

When mentioning that aspect of the present invention relates to the crystal formation of compound 2f, degree of crystallinity is suitably greater than about 60%, more suitably greater than about 80%, be preferably greater than about 90%, more preferably greater than about 95%.Most preferably degree of crystallinity is greater than about 98%.

The compound 2f of crystal form A has and the essentially identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 5, and has 10 main peaks shown in the table 3 (2-θ angle value) substantially.Should be appreciated that the 2-θ angle value for different instruments or different sample X-ray powder diffraction pattern can change slightly, thus aforementioned value can not to be interpreted as be absolute.

As everyone knows, the X-ray powder diffraction pattern that (for example instrument of Shi Yonging or equipment) can obtain having one or more measuring error under different measuring conditions.Particularly, known usually, the intensity of X-ray powder diffraction pattern may fluctuate under different measuring conditions.Therefore, be to be understood that, the compound 2f of crystal form A of the present invention is not limited to have the crystal with the identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 5, and any have with the crystal of the essentially identical X-ray powder diffraction pattern of X-ray powder diffraction pattern shown in Figure 5 all within the scope of the invention.The technician in X-ray powder diffraction field can judge the basically identical of X-ray powder diffraction pattern.

The technician in X-ray powder diffraction field will be appreciated that the relative intensity at peak may be subjected to, for example, the particle more than 30 microns and the influence of non-single length-to-diameter ratio, this may influence the analysis of sample.Those skilled in the art will be appreciated that also reflection position may be subjected to the definite height of sample in diffractometer and the influence of diffractometer zero correction.The profile pattern of sample also has small influence.Therefore, given diffractogram data can not be used as absolute figure.(Jenkins, R and Snyder, R.L. ' Introduction to X-Ray Powder Diffractometry ' John Wiley ﹠amp; Sons 1996; Bunn, C.W. (1948), Chemical Crystallography, Clarendon Press, London; Klug, H.P. and Alexander, L.E. (1974), X-Ray Diffraction Procedures).

Usually, the measuring error of the diffraction angle in the X-ray powder diffraction pattern is ± 0.1 °, the X-ray powder diffraction pattern in considering Fig. 1,3 and 5, and understand table 1,2 and at 3 o'clock, should consider the measuring error of this degree.In addition, be to be understood that intensity may fluctuate with experiment condition and specimen preparation (preferred orientation).

Used ins and outs

X-ray diffraction

Table 4

* relative intensity derives from the diffractogram that uses fixed slit to record

Analytical instrument: PANalytical Cubix PRO

Place on the silicon single crystal wafer bearing (zero background support) and sample is spread out lamellar measure x-ray diffraction pattern by sample crystalline salt.Rotary sample to be improving counting statistics, and uses the x-ray irradiation sample, and described X-ray is focused on pipe and produced under the condition of 45kV and 40mA by length-thin copper, and it has

Wavelength.Calibrated X-ray source process is arranged on the automated variable divergent slit of V20, and reflected ray directly passes the anti-scatter slit of 2mm and the detection slit of 0.2mm.Under θ-θ pattern, make per 0.02 ° 2-θ increment (counting scan pattern) exposure of sample 100 seconds in 2 ° to 40 ° the 2-θ scope.The technician in X-ray powder diffraction field will be appreciated that the relative intensity at peak may be subjected to, for example, the particle more than 30 microns and the influence of non-single length-to-diameter ratio, this may influence the analysis of sample.Those skilled in the art will be appreciated that also reflection position may be subjected to the definite height of sample in diffractometer and the influence of diffractometer zero correction.The planarization of sample also has small influence.Therefore, the diffractogram data of proposition can not be used as absolute figure.

The differential scanning calorimetric

Analytical instrument: TA Instruments Q1000.

Typically, the material that the is lower than 5mg constant heating rate with 10 ℃/minute in 25 ℃-300 ℃/25 ℃-180 ℃ temperature range that is contained in the aluminium crucible with perforation lid is heated.Make

With nitrogen as sweep gas-flow 50ml/ minute.

Embodiment

The general experimental technique that is used for embodiment 1-3

Preparation HPLC

Instrument: the Waters ZMD LC-MS NO.LD352 of system operates under the electro-spray ionization pattern.

Mobile phase A: 0.1% aqueous formic acid

Mobile phase B: 0.1% formic acid acetonitrile solution

Post: Genesis C18 4 μ m 50 * 4.6mm

Gradient:

Time (minute)	％B
		0	5
7	95
		9	95
9.5	5
		13	5

Flow: 1.0ml/ minute.

PDA sweep limit: 210-400nm.

Long method (Long method)

Instrument: the Waters ZQ LC-MS NO.LAA of system 254 operates under the electro-spray ionization pattern.

Mobile phase A: 0.1% aqueous formic acid

Mobile phase B: 0.1% formic acid acetonitrile solution

Post: Genesis C18 4 μ m 50 * 4.6mm

Gradient:

Time (minute)

％B

0	5
		20	95
23	95
		24	5
25	5

Flow: 2.0ml/ minute.

PDA sweep limit: 210-400nm.

Embodiment 1

(a) storehouse synthetic (2a-j)

To 2-fluoro-5-(4-oxygen-3,4-dihydro-phthalazines-1-ylmethyl)-phenylformic acid (1) (29mg, add in DMA 0.1mmol) (0.5ml) solution DIPEA (0.02 μ L, 0.11mmol), HBTU (42mg, 0.10mmol), add suitable piperidine derivative (0.10mmol) then.At room temperature stirring reaction is 16 hours.Then with rough sample preparation HPLC purifying.

*=long method

Embodiment 2

(a) storehouse synthetic (4a-c)

To 3-(5,8-two fluoro-4-oxygen-3,4-dihydro-phthalazines-1-ylmethyl)-phenylformic acid (3) (32mg, 0.1mmol) DMA (0.5ml) solution in add DIPEA (0.02 μ L, 0.11mmol), (42mg 0.10mmol), adds suitable piperidine derivative (0.10mmol) to HBTU then.At room temperature stirring reaction is 16 hours.Then with rough sample preparation HPLC purifying.

Embodiment 3

(a) storehouse synthetic (6a-c)

To 4-(4-oxygen-3,4-dihydro-phthalazines-1-ylmethyl)-pyridine-2-carboxylic acids (5) (28mg, add in DMA 0.1mmol) (0.5ml) solution DIPEA (0.02 μ L, 0.11mmol), HBTU (42mg, 0.10mmol), add suitable piperidine derivative (0.10mmol) then.At room temperature stirring reaction is 16 hours.Then with rough sample preparation HPLC purifying.

* long method

+ following analysis mode LC-MS method

The general experimental technique that is used for embodiment 4-45

Analysis mode LC-MS

The LC-MS data derive from following system, the HPLC part comprises Agilent1100 usually in the described system, Waters Alliance HT (2790 and 2795) device or HP 1100 pumps and the diode array that has the CTC automatic sampler, and at Phenomenex Gemini C18 5mm, 50 * 2mm post (or similar) is gone up operation, with acid elutriant (for example, in 4 minutes, make water/acetonitrile 0-95% gradient elution, described water/acetonitrile elutriant contains 5% formic acid solution, and described formic acid solution is 1% formic acid at water: acetonitrile (v/v) is the solution in 50: 50 the mixture; Or the solvent system with methyl alcohol replacement acetonitrile of use equivalent), or alkaline eluant (for example, in 4 minutes, make water/acetonitrile 0-95% gradient elution, described water/acetonitrile elutriant contains 5% 880 ammonia solutions, and described 880 ammonia solutions are the solution of 0.1%880 ammonia in the acetonitrile mixture) wash-out; The MS part comprises the Waters ZQ mass spectrograph of scanning appropriate mass scope usually.Generate positive and negative base peak intensity of electron spray(ES) (ESI) chromatogram and 220-300nmUV hypersorption chromatogram, and provide the m/z value; Usually, only provide the ion of expression parent nucleus quality, except as otherwise noted, fiducial value (M+H) ⁺Be positive ion mode, (M-H)-be negative ion mode.

The NMR spectrum

The NMR data that provide are to use, and for example, Bruker DPX-400 spectrometer is measured under 400MHz, and the NMR data are δ value form, for main proton to be measured, are unit with PPM (ppm).Except as otherwise noted, used solvent is CDCl ₃(using tetramethylsilane (TMS)) or DMSO-d as interior mark ₆Use following initialism: s, unimodal; D, bimodal; T, triplet; Q, quartet; M, multiplet; Br, broad peak.

Embodiment 4

4-(4-fluoro-3-(4-(2-methoxy ethoxy) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (7)

With 4-(2 methoxy ethoxy) piperidine hydrochlorate (103mg, 0.53mmol) and triethylamine (0.210ml, 1.51mmol) processing 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (150mg, 0.50mmol) N,N-dimethylacetamide (4ml) solution.Add O-benzotriazole-1-base-N, N, N ', (253mg 0.67mmol), and at room temperature stirred the solution that obtains 4.5 hours N '-tetramethyl-urea hexafluorophosphate.Filter rough reaction mixture, use preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying then, the water that reduces gradually with polarity (contains 1% NH ₃) and the MeCN mixture as elutriant.The fraction that will contain required compound is evaporated to dry also freeze-drying and obtains gelatinoid, described gelatinoid is placed a small amount of diethyl ether and methylene dichloride and makes its evaporation, 55 ℃ of following vacuum-dryings 2 hours, obtain required compound then, be white solid (151mg, productive rate 68.3%); ¹H NMR (400.132MHz, DMSO) δ 1.28-1.36 (1H, m), 1.40-1.49 (1H, m), 1.68-1.75 (1H, m), 1.82-1.90 (1H, m), 2.99-3.06 (1H, m), 3.25 (3H, s), 3.26-3.32 (2H, m), 3.44 (2H, t), 3.53-3.58 (3H, m), 3.90-3.98 (1H, m), 4.33 (2H, s), 7.21 (1H, t), 7.33-7.35 (1H, m), 7.39-7.43 (1H, m), 7.81-7.91 (2H, m), 7.97 (1H, d), 8.27 (1H, dd), 12.56 (1H, s); M/z (LC-MS, ESI+), RT=1.65 (M+H 440.6).

Embodiment 5

Multiple parallel synthesizes (8a-f)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) phenylformic acid (1) (120mg methyl), 0.40mmol), triethylamine (144 μ l, 1.03mmol) and O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (200mg, 0.53mmol) N,N-dimethylacetamide (3ml) solution join in the suitable piperidines (0.50mmol), and mixture at room temperature stirred spends the night.Use the syringe-type strainer filter reaction mixture of 0.45 μ m then, and directly use preparation HPLC (WatersXBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying filtrate, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required product merges, and is evaporated to dry and freeze-drying, obtains final product, and final product is dissolved in several methylene dichloride and the diethyl ether (1-2ml).With solvent evaporation, 60 ℃ of following vacuum-drying resistatess 3 hours, obtain required compound then.

8a:-4-[[4-fluoro-3-[4-(3-methoxyl group phenoxy group) piperidines-1-carbonyl] phenyl] methyl]-2H-phthalazines-1-ketone; ¹H NMR (400.132MHz, DMSO) δ 1.46-1.67 (2H, m), 1.80-1.87 (1H, m), 1.95-2.03 (1H, m), and 3.13-3.20 (1H, m), 3.33-3.50 (2H, m), 3.73 (3H, s), 3.92-4.01 (1H, m), 4.33 (2H, s), 4.60-4.66 (1H, m), and 6.51-6.58 (3H, m), 7.16-7.25 (2H, m), 7.36-7.44 (2H, m), 7.80-7.90 (2H, m), 7.98 (1H, d), 8.26 (1H, dd), 12.57 (1H, s).

8b:-4-[[4-fluoro-3-[4-(4-methoxyl group phenoxy group) piperidines-1-carbonyl] phenyl] methyl]-2H-phthalazines-1-ketone; ¹H NMR (400.132MHz, DMSO) δ 1.44-1.65 (2H, m), 1.76-1.84 (1H, m), 1.91-1.99 (1H, m), and 3.10-3.17 (1H, m), 3.32-3.49 (2H, m), 3.70 (3H, s), 3.92-3.99 (1H, m), 4.33 (2H, s), 4.45-4.51 (1H, m), 6.83-6.94 (4H, m), 7.22 (1H, t), 7.35-7.44 (2H, m), 7.80-7.90 (2H, m), 7.97 (1H, d), 8.26 (1H, dd), 12.57 (1H, s).

8c:-4-[[4-fluoro-3-(4-propoxy-piperidines-1-carbonyl) phenyl] methyl]-2H-phthalazines-1-ketone; ¹H NMR (400.132MHz, DMSO) δ 0.87 (3H, t), 1.27-1.36 (1H, m), and 1.39-1.55 (3H, m), 1.66-1.74 (1H, m), and 1.82-1.89 (1H, m), 2.99-3.07 (1H, m), and 3.24-3.29 (2H, m), 3.36 (2H, td), and 3.48-3.54 (1H, m), 3.89-3.98 (1H, m), 4.33 (2H, s), 7.21 (1H, t), and 7.33-7.35 (1H, m), 7.39-7.43 (1H, m), and 7.81-7.91 (2H, m), 7.97 (1H, d), 8.27 (1H, dd), 12.56 (1H, s).

8d:-4-[[4-fluoro-3-[4-(2-methoxyl group phenoxy group) piperidines-1-carbonyl] phenyl] methyl]-2H-phthalazines-1-ketone; ¹H NMR (400.132MHz, DMSO) δ 1.49-1.68 (2H, m), 1.76-1.84 (1H, m), 1.91-1.99 (1H, m), and 3.10-3.17 (1H, m), 3.35-3.50 (2H, m), 3.76 (3H, s), 3.93-4.00 (1H, m), 4.34 (2H, s), 4.47-4.53 (1H, m), 6.85-7.05 (4H, m), 7.22 (1H, t), 7.36-7.43 (2H, m), 7.80-7.90 (2H, m), 7.98 (1H, d), 8.26 (1H, dd), 12.57 (1H, s).

8e:-4-[[4-fluoro-3-(4-phenyl methoxyl group piperidines-1-carbonyl) phenyl] methyl]-2H-phthalazines-1-ketone; ¹H NMR (400.132MHz, DMSO) δ 1.37-1.59 (2H, m), 1.72-1.79 (1H, m), 1.88-1.94 (1H, m), and 3.01-3.08 (1H, m), 3.27-3.38 (2H, m), 3.63-3.68 (1H, m), 3.90-3.98 (1H, m), 4.33 (2H, s), 4.52 (2H, s), 7.21 (1H, t), 7.26-7.31 (1H, m), 7.33-7.38 (5H, m), and 7.39-7.43 (1H, m), 7.81-7.91 (2H, m), 7.97 (1H, d), 8.27 (1H, dd), 12.56 (1H, s).

8f:-2-[4-[1-[2-fluoro-5-[(4-oxygen-3H-phthalazines-1-yl) methyl] benzoyl] piperidin-4-yl] oxygen base phenyl] acetonitrile; ¹H NMR (399.902MHz, DMSO) δ 1.47-1.68 (2H, m), 1.81-1.88 (1H, m), 1.96-2.04 (1H, m), and 3.13-3.21 (1H, m), 3.32-3.40 (1H, m), 3.43-3.51 (1H, m), 3.92-4.03 (3H, m), 4.34 (2H, s), 4.62-4.68 (1H, m), and 7.00-7.03 (2H, m), 7.21-7.28 (3H, m), 7.37-7.44 (2H, m), 7.80-7.90 (2H, m), 7.98 (1H, d), 8.27 (1H, dd), 12.58 (1H, s).

[initial piperidines :-CAS 1000516-48-2]

Embodiment 6

4-(4-fluoro-3-(3H-spiral shell [isobenzofuran-1,4 '-piperidines]-1 '-Ji carbonyl) benzyl) phthalazines-1 (2H)-ketone (9)

With triethylamine (0.168mg, 1.21mmol) and O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (256mg, 0.68mmol) processing 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) (144mg is 0.48mmol) with 3H-spiral shell [isobenzofuran-1,4 '-piperidines] hydrochloride (109mg for phenylformic acid (1), 0.48mmol) N,N-dimethylacetamide (2ml) solution.The mixture that obtains was at room temperature stirred 5 hours, filter then, and with preparation HPLC (Waters XBridge PrepC18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 0.1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to dry also freeze-drying and obtains required compound, is white solid (118mg, productive rate 52.1%); ¹HNMR (400.132MHz, DMSO) δ 1.53-1.62 (2H, m), 1.73-1.83 (3H, m), 1.91-1.99 (1H, m), 3.10-3.18 (1H, m), 4.40 (2H, s), and 4.54-4.60 (1H, m), 5.03-5.11 (2H, m), 7.26-7.37 (5H, m), 7.45-7.50 (2H, m), 7.83-7.93 (2H, m), 8.02 (1H, d), 8.30 (1H, d), 12.13-12.59 (1H, br s); M/z (LC-MS, ESI+), RT=2.14 (M+H 470.9).

Embodiment 7

(a) 4-(4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (2a)

With the 4-hydroxy piperidine (396mg, 3.92mmol) and triethylamine (1ml 7.17mmol) handles 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (1g, N,N-dimethylacetamide 3.35mmol) (30ml) solution.Add O-benzotriazole-1-base-N, N, N ', (1.77g 4.67mmol), at room temperature stirred the solution that obtains 17 hours N '-tetramethyl-urea hexafluorophosphate.Then reaction mixture is poured in the water (300ml) also with methylene dichloride (2 * 200ml) extractions.The organic extract that merges is cleaned with salt solution, use dried over mgso, filtration and evaporation obtain raw product, and with quick silicon-dioxide chromatogram purification raw product, gradient is the dichloromethane solution of 0-20%MeOH.Pure fraction is evaporated to drying, obtains required compound, be light yellow gluey material (1.24g, productive rate 97%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.42-1.54 (1H, m), 1.55-1.67 (2H, m), 1.76-1.84 (1H, m), and 1.92-2.01 (1H, m), 3.05-3.15 (1H, m), 3.38-3.55 (2H, m), 3.94-3.99 (1H, m), 4.14-4.22 (1H, m), 4.27 (2H, s), 7.02 (1H, t), 7.26-7.32 (2H, m), 7.70-7.79 (3H, m), 8.44-8.48 (1H, m), 10.07 (1H, s); M/z (LC-MS, ESI+), RT=1.44 (M+H 382.1).

(b) methyl 4-[1-[2-fluoro-5-[(4-oxygen-3H-phthalazines-1-yl)] benzoyl] piperidin-4-yl] oxygen base benzonitrile (10)

With polymkeric substance loading type triphenylphosphine (ex-Biotage, 742mg, 1.37mmol) and azo-2-carboxylic acid's di tert butyl carbonate (300mg, 1.30mmol) processing 4-hydroxy phenyl nitrile (48mg, 0.40mmol) and 4-(4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (2a) (150mg, methylene dichloride 0.39mmol) (5ml) solution.Reaction mixture was at room temperature shaken 4-5 hour.Then mixture is removed by filter resin, and use the washed with methanol filter cake.Filtrate is evaporated, obtain wax shape yellow solid, with multiple preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, earlier water (containing 0.1%TFA) that reduces gradually with polarity and MeCN are as elutriant, and water that reduces gradually with polarity (containing 1% ammonia) and MeCN are as elutriant (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide then, diameter 19mm, long 100mm).The fraction that will contain required compound is evaporated to dry and freeze-drying, obtains required product (17mg, productive rate 8%); ¹H NMR (400.132MHz, DMSO) δ 1.49-1.69 (2H, m), 1.83-1.91 (1H, m), 1.99-2.06 (1H, m), and 3.14-3.22 (1H, m), 3.32-3.51 (2H, m), 3.94-4.04 (1H, m), 4.33 (2H, s), 4.78-4.84 (1H, m), 7.15-7.18 (2H, m), 7.23 (1H, t), 7.36-7.44 (2H, m), 7.75-7.78 (2H, m), 7.80-7.90 (2H, m), 7.97 (1H, d), 8.26 (1H, d), 12.56 (1H, s); M/z (LC-MS, ESI+), RT=2.20 (M+H 483.6).

Embodiment 8

4-(4-fluoro-3-(4-isopropoxy piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (11)

Under 25 ℃, with 4-isopropoxy piperidine hydrochlorate (120mg, 0.67mmol) and triethylamine (0.206ml, 1.48mmol) N, disposable 2-fluoro-5-((the 4-oxygen-3 that is stirring that joins of N dimethyl formamide (2ml) solution, 4-dihydro phthalazines-1-yl) phenylformic acid (1) (200mg methyl), 0.67mmol), triethylamine (0.206ml, 1.48mmol) and O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (381mg, 1.01mmol) N, in N dimethyl formamide (2ml) solution.The solution that obtains was stirred 4 hours down at 25 ℃.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 30mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (172mg, productive rate 60.6%); ¹H NMR (399.902MHz, DMSO) δ 1.06-1.11 (6H, m), 1.21-1.46 (2H, m), 1.61-1.87 (2H, m), 3.04 (1H, t), 3.62 (2H, td), 3.70 (1H, quintet), 3.92-4.01 (1H, m), 4.07 (1H, q), 4.33 (2H, s), and 7.19-7.25 (1H, m), 7.32-7.37 (1H, m), 7.38-7.44 (1H, m), 7.81-7.92 (2H, m), 7.96-8.00 (1H, m), 8.26-8.29 (1H, m), 12.58 (1H, s); M/z (LC-MS, ESI+), RT=1.98 (M+H 424.6).

Embodiment 9

Under 25 ℃, with 4-isopropoxy piperidine hydrochlorate (128mg, 0.71mmol) and triethylamine (0.219ml, N 1.57mmol), disposable 3-((the 4-oxygen-3 that is stirring that joins of N dimethyl formamide (2ml) solution, 4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1b) (200mg, 0.71mmol), triethylamine (0.219ml, 1.57mmol) and O-benzotriazole-1-base-N, N, N ', (406mg is in DMF 1.07mmol) (2ml) solution for N '-tetramethyl-urea hexafluorophosphate.The solution that obtains was stirred 4 hours down at 25 ℃.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 30mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (172mg, productive rate 59.4%); ¹H NMR (399.902MHz, DMSO) δ 1.06-1.10 (6H, m), 1.21-1.46 (2H, m), 1.57-1.90 (2H, m), 3.00-3.13 (2H, m), 3.55-3.64 (2H, m), and 3.64-3.75 (1H, m), 3.88-4.02 (1H, m), 4.33-4.38 (2H, m), 7.21 (1H, dt), 7.31-7.42 (3H, m), 7.85 (2H, m), 7.96 (1H, m), 8.27 (1H, m), 12.59 (1H, s); M/z (LC-MS, ESI+), RT=1.89 (M+H 406.6).

Embodiment 10

(a) 4-cyclobutoxy group pyridine (13)

With pyridine-4-alcohol (6.0g, 63.09mmol), cyclobutanol (5.00g, 69.40mmol) and triphenylphosphine (18.20g 69.40mmol) joins among the THF (250ml), and stirs 10 minutes.Slow adding diisopropyl azo-2-carboxylic acid in this solution (13.49ml, 69.40mmol).To be reflected at 50 ℃ when adding finishes immediately stirred 1 hour down.Solvent evaporation also is dissolved in diethyl ether with rough resistates.In this solution, add a small amount of triphenylphosphine oxidation thing, and will react and stir 30 minutes, obtain solid, this solid filtering is fallen.With solvent evaporation, and with the lurid gelatinoid of 2.0M HCl acidifying, (1 * 75ml) extraction is then with the solid potassium hydroxide aqueous solution that alkalizes with diethyl ether.(3 * 75ml) extract this solution, pass through MgSO to use diethyl ether then ₄Dry organic layer filters and evaporation obtains yellow jelly matter.Be purified by under 0.60mBar, distilling, be collected in 80 ℃ of distillatory cuts, obtain required compound, be colourless oily matter (4.70g, productive rate 49.9); ¹H NMR (400.132MHz, CDCl ₃) δ 1.78-1.66 (1H, m), 1.95-1.86 (1H, m), 2.24-2.14 (2H, m), 2.51-2.43 (2H, m), 4.69 (1H, quintet), 6.70 (2H, d), 8.40 (2H, d); M/z (LC-MS, ESI+), RT=1.76 (M+H 150).

(b) 4-cyclobutoxy group piperidines (14)

In the nitrogen atmosphere of 5bar, under 25 ℃ with 4-cyclobutoxy group pyridine (13) (3.8g, 25.47mmol) and 5% rhodium aluminum oxide (0.38g, EtOH 0.05mmol) (50ml) solution stirring 16 hours.GCMS analyze to show and not react, and therefore with mixture heating up to 60 ℃, 5 hours, only shows that still micro-product forms but analyze.Add platinum oxide (0.2g) catalyzer, and under 60 ℃, 5bar, stirs and spend the night, but still do not have more product formation.Reaction mixture is moved in the autoclave, and under 80 ℃, 50bar heated overnight, do not observe more product and form, therefore temperature is increased to 100 ℃, pressure is increased to 80bar, placement is spent the night, and analyzes still to show do not have more product to form.Therefore add 5% rhodium carbon (0.2g) and heated overnight under 120 ℃, 100bar.Analyze and show there is not residual raw material.To react cooling and filtration,, obtain yellow oil solvent evaporation.Come the purification of crude product by distillation under 0.55mBar, be collected in 55 ℃ of distillatory cuts, obtaining product is colorless oil, and this product is used for next step, and need not to be further purified.

(c) 4-(3-(4-cyclobutoxy group piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (15)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1b) (0.20g, 0.67mmol) and O-benzotriazole-1-base-N, N, N ', (0.381g 1.01mmol) joins N to N '-tetramethyl-urea hexafluorophosphate, in the dinethylformamide (30ml), and in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.179ml, 1.01mmol), add then 4-cyclobutoxy group piperidines (14) (0.104g, 0.67mmol).To react and stir 2 hours, and use preparation HPLC (Waters XBridge Prep C18OBD post, 5 μ silicon-dioxide, diameter 21mm, long 100mm) purifying then, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white solid (0.017g, 5.82%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.56-1.44 (2H, m), 1.72-1.64 (2H, m), 2.00-1.84 (4H, m), and 2.23-2.14 (2H, m), 3.14-3.04 (1H, m), 3.48-3.41 (2H, m), 3.58-3.52 (1H, m), 4.00 (1H, quintet), 4.10-4.03 (1H, m), 4.26 (2H, s), 7.01 (1H, t), 7.31-7.24 (2H, m), 7.77-7.70 (3H, m), 8.47-8.44 (1H, m), 9.90 (1H, s); M/z (LC-MS, ESI+), RT=2.15 (M+H 436).

Embodiment 11

With 2-fluoro-5-((4-oxygen-3,4-dihydro-phthalazines-1-yl) methyl)-phenylformic acid (1) (0.2g, 0.67mmol), 4-(allyloxy) piperidines (0.095g, 0.67mmol) and O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (0.381g, 1.01mmol) join N, in the dinethylformamide (30ml).In this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.179ml, 1.01mmol), and will react and stir 2 hours, use preparation HPLC (Waters XBridge Prep C18 OBD post then, 5 μ silicon-dioxide, diameter 21mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (0.225g, 80%); ¹H NMR (400.132MHz, DMSO) δ 1.37-1.31 (1H, m), 1.51-1.42 (1H, m), 1.74-1.70 (1H, m), 1.90-1.86 (1H, m), 3.07-3.00 (1H, m), and 3.35-3.26 (2H, m), 3.60-3.54 (1H, m), 3.94-3.90 (1H, m), 3.98 (2H, d), 4.33 (2H, s), 5.13 (1H, dq), 5.26 (1H, dq), 5.94-5.85 (1H, m), 7.21 (1H, t), 7.35-7.33 (1H, m), 7.43-7.39 (1H, m), 7.83 (1H, t), 7.89 (1H, t), 7.97 (1H, d), 8.27 (1H, d), 12.58 (1H, s); M/z (LC-MS, ESI+), RT=1.93 (M+H 422).

Embodiment 12

(a) 4-(difluoro-methoxy) piperidines-1-formaldehyde (17)

(10.8g 83.62mmol) is dissolved in tetrahydrofuran (THF) (150ml) with 4-hydroxy piperidine-1-formaldehyde.(5.22g 108.70mmol), and will react and stir 30 minutes to add sodium hydride in this solution.(9.21g 41.81mmol) also will react further stirring 30 minutes, and (8.68g 100.34mmol), will react after the adding and stir 30 minutes slowly to add chlorodifluoromethane then to add 15-hat-5.Finish reaction mixture and finish reaction with saturated brine (75ml), (MgSO is used in 3 * 100ml) extractions with diethyl ether ₄Dry organic layer filters and evaporation, obtains yellow liquid.Come the purification of crude product by distillation under 0.8mBar, be collected in 80 ℃ of distillatory cuts, obtain desired substance, be colourless liquid (2.70g, 18.02%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.93-1.70 (4H, m), 3.32-3.26 (1H, m), 3.60-3.47 (2H, m), 3.74-3.67 (1H, m), 4.50 (1H, septet), 6.28 (1H, t), 8.03 (1H, s).

(b) 4-(difluoro-methoxy) piperidines (18)

With 4-(difluoro-methoxy) piperidines-1-formaldehyde (17) (2.7g, 15.07mmol) and potassium hydroxide (2.96g, 52.74mmol) water-soluble (40ml), and vigorous stirring 3 hours.(MgSO is used in 3 * 75ml) extractions with dme ₄Dry organic layer filters and evaporation, obtains orange liquid.Come the purification of crude product by distillation under 0.70mBar, be collected in 25 ℃ of distillatory cuts, obtain desired substance, be colorless oil (1.100g, 48.3%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.50 (1H, s), 1.66-1.57 (2H, m), 1.95-1.89 (2H, m), 2.70-2.64 (2H, m), 3.09 (2H, dt), 4.25-4.19 (1H, m), 6.24 (1H, t).

(c) 4-(3-(4-(difluoro-methoxy) piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (19)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1) (0.2g, 0.67mmol), 4-(difluoro-methoxy) piperidines (18) (0.101g, 0.67mmol) and O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (0.381g, 1.01mmol) join N, in the dinethylformamide (30ml).Add N-ethyl-N-sec.-propyl third-2-amine (0.179ml to this solution, 1.01mmol), and will react and stir 2 hours, use preparation HPLC (Waters XBridge Prep C18 OBD post then, 5 μ silicon-dioxide, diameter 21mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white solid (0.266g, 92%); ¹H NMR (400.132MHz, DMSO) δ 1.53-1.45 (1H, m), 1.65-1.56 (1H, m), 1.83-1.74 (1H, m), 1.98-1.92 (1H, m), 3.15-3.09 (1H, m), 3.40-3.26 (2H, m), 4.01-3.91 (1H, m), 4.33 (2H, s), 4.41-4.35 (1H, m), 6.75 (1H, t), 7.22 (1H, t), 7.38-7.36 (1H, m), 7.44-7.40 (1H, m), 7.83 (1H, t), 7.88 (1H, t), 7.97 (1H, d), 8.27 (1H, d), 12.58 (1H, s); M/z (LC-MS, ESI+), RT=1.98 (M+H 432).

Embodiment 13

4-[[4-fluoro-3-[4-(tetrahydrofuran (THF)-2-ylmethoxy) piperidines-1-carbonyl] phenyl] methyl]-2H-phthalazines-1-ketone (20)

Under 25 ℃, air atmosphere, with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (509mg, 1.34mmol) disposable 2-fluoro-5-((the 4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) phenylformic acid (200mg methyl), 0.67mmol) and triethylamine (0.206ml, N 1.48mmol) is in dinethylformamide (2.5ml) solution.The solution that obtains was stirred 10 minutes down at 25 ℃.Dropwise add then 4-((tetrahydrofuran (THF)-2-yl) methoxyl group) piperidine hydrochlorate (164mg, 0.74mmol) and triethylamine (dinethylformamide (1ml) solution stirs the solution that obtains 10 hours down at 25 ℃ for 0.206ml, N 1.48mmol).With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 30mm, long 100mm) purification of crude mixture, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is solid (238mg, 76%); ¹H NMR (400.132MHz, DMSO) δ 1.01 (2H, dt), 1.31-1.37 (1H, m), 1.43-1.50 (1H, m), 1.51-1.58 (1H, m), 1.68-1.73 (1H, m), and 1.74-1.93 (4H, m), 3.00-3.07 (1H, m), 3.17-3.18 (1H, m), 3.39 (2H, d), 3.53-3.59 (1H, m), 3.70-3.75 (1H, m), 3.90 (2H, ddd), 4.33 (2H, s), 7.21 (1H, t), 7.33-7.35 (1H, m), 7.39-7.43 (1H, m), 7.83 (1H, td), 7.89 (1H, td), 7.97 (1H, d), 8.27 (1H, dd), 12.56 (1H, s) m/z (LC-MS, ESI+), RT=1.71 (M+H466.5).

Embodiment 14

(a) 4-hydroxy-4-methyl piperidines-1-carboxylic acid tert-butyl ester (21)

Under 25 ℃, air atmosphere, with sodium hydride (60% dispersion in mineral oil) (158mg, 3.96mmol) in 1 minute time, join 4-methyl piperidine-4-alcohol hydrochloride (500mg in batches, 3.30mmol) and triethylamine (0.506ml, 3.63mmol) N, in dinethylformamide (8ml) solution.The suspension that obtains was stirred 20 minutes at 25 ℃.Disposable then adding tert-Butyl dicarbonate (1.668ml, DMF 7.25mmol) (2ml) solution, and the suspension that obtains stirred 18 hours at 25 ℃.Water (125ml) and saturated brine (25ml) diluted reaction mixture, (3 * 50ml) clean to use methylene dichloride then successively.Pass through MgSO ₄Dry organic layer filters and evaporation, obtains rough required product, and it need not to be further purified in use; 1H NMR (400.132MHz, CDCl3) δ 1.46 (9H, s), 1.51-1.56 (7H, m), 3.21-3.28 (2H, m), 3.65-3.73 (2H, m)

(b) 4-methoxyl group-4-methyl piperidine-1-carboxylic acid tert-butyl ester (22)

Under room temperature, air atmosphere, with sodium hydride (60% dispersion in mineral oil) (198mg, 4.95mmol) join 4-hydroxy-4-methyl piperidines-1-carboxylic acid tert-butyl ester (21) in 30 seconds time (710mg, N 3.30mmol) is in dinethylformamide (12ml) solution in batches.The suspension that obtains was stirred 20 minutes at 25 ℃.(0.411ml 6.60mmol), and stirs the suspension that obtains 18 hours at 25 ℃ to add methyl iodide then.Water (150ml) and saturated brine (25ml) diluted reaction mixture, (3 * 100ml) clean to use methylene dichloride then successively.Organic layer passes through MgSO ₄Drying is filtered and evaporation, obtains rough required product, and it directly uses in next step.

(c) 4-methoxyl group-4-methyl piperidine (23)

Under 25 ℃, air atmosphere, (2.54ml, (756mg is in methylene dichloride 3.30mmol) (10ml) solution 32.97mmol) dropwise to join 4-methoxyl group-4-methyl piperidine-1-carboxylic acid tert-butyl ester (22) with trifluoroacetic acid.The solution that obtains was stirred 15 hours down at 25 ℃.The crude product ion-exchange chromatogram purification uses the SCX post.Use 7M NH ₃The required product of/MeOH wash-out from post, and pure fraction is evaporated to drying, obtain required compound, be oily matter (235mg, 55.2%); ¹H NMR (400.132MHz, CDCl3) δ 1.15 (3H, s), 1.41-1.49 (2H, m), 1.68-1.74 (2H, m), 2.76 (2H, dt), 2.93 (2H, dt), 3.19 (3H, s), 3.49 (1H, s).

(d) 4-[[4-fluoro-3-(4-methoxyl group-4-methyl piperidine-1-carbonyl) phenyl] methyl]-2H-phthalazines-1-ketone (24)

Under room temperature, air atmosphere, with O-(1H-benzotriazole-1-yl)-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (0.597g, 1.57mmol) disposable 2-fluoro-5-((the 4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) phenylformic acid (1) (0.188g methyl), 0.63mmol) and triethylamine (0.193ml, N 1.39mmol) is in dinethylformamide (2ml) solution.The solution that obtains was stirred 15 minutes down at 25 ℃.(dinethylformamide (1ml) solution stirs the solution that obtains 4 hours down at 25 ℃ then for 89.5mg, N 0.69mmol) dropwise to add 4-methoxyl group-4-methyl piperidine (23).With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 21mm, long 100mm) purification of crude mixture, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is solid (0.110g, 42.7%); ¹H NMR (400.132MHz, DMSO) δ 1.11 (3H, s), 1.25-1.34 (1H, m), 1.42 (1H, td), 1.56 (1H, d), 1.75 (1H, dd), 3.10-3.18 (4H, m), 4.09 (3H, q), 4.32 (2H, s), 7.21 (1H, t), 7.31 (1H, dd), 7.39-7.43 (1H, m), 7.83 (1H, td), 7.88 (1H, td), 7.96 (1H, dd), 8.27 (1H, dd), 12.57 (1H, s); M/z (LC-MS, ESI+), RT=1.79 (M+H409).

Embodiment 15

4-[[2-(4-third-2-base oxygen phenylpiperidines-1-carbonyl) pyridin-4-yl] methyl]-2H-phthalazines-1-ketone (25)

Under 25 ℃, with 4-isopropoxy piperidine hydrochlorate (192mg, 1.07mmol) and triethylamine (0.327ml, N 2.35mmol), disposable 4-((the 4-oxygen-3 that is stirring that joins of dinethylformamide (3ml) solution, 4-dihydro phthalazines-1-yl) methyl)-pyridine carboxylic acid (5) (300mg, 1.07mmol), triethylamine (0.327ml, 2.35mmol) and O-benzotriazole-1-base-N, N, N ', (607mg is in DMF 1.60mmol) (3ml) solution for N '-tetramethyl-urea hexafluorophosphate.The solution that obtains was stirred 4 hours down at 25 ℃.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 30mm, long 100mm) purification of crude mixture, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to dry and freeze-drying, obtains required compound, is solid (257mg, 59.3%); ¹H NMR (400.132MHz, DMSO) δ 1.08 (6H, dd), 1.28-1.46 (2H, m), 1.65-1.74 (1H, m), 1.78-1.87 (1H, m), 3.12 (1H, ddd), 3.23-3.31 (1H, m), 3.42-3.51 (1H, m), 3.62 (1H, m), 3.69 (1H, quintet), 3.91-4.00 (1H, m), 4.40 (2H, s), 7.39 (1H, dd), 7.49 (1H, d), 7.83-7.97 (3H, m), 8.28 (1H, dd), 8.46-8.49 (1H, m), 12.60 (1H, s); M/z (LC-MS, ESI+), RT=1.54 (M+H 407).

Embodiment 16

(a) 4-(4-fluoro-3-(4-(pyrimidine-4-base oxygen base) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (26a)

At 20 ℃, (125mg 1.30mmol) disposablely joins 4-(4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) benzyl) (150mg is in tetrahydrofuran (THF) 0.39mmol) (2ml) solution for phthalazines-1 (2H)-ketone (2a) with sodium tert-butoxide.The suspension that obtains was stirred 10 minutes down at 20 ℃.(49.5mg 0.43mmol), and down stirs mixture 2 hours at 50 ℃ disposable adding 4-chloropyrimide hydrochloride.(41mg, 0.43mmol), (65mg 0.43mmol), heats mixture 2 hours down at 50 ℃ disposable adding sodium tert-butoxide, at room temperature stirs then and spends the night to add 4-chloropyrimide hydrochloride then.The raw product ion-exchange chromatogram purification uses the SCX post.Use 2M NH ₃The required product of/MeOH wash-out from post, and pure fraction is evaporated to drying, obtain raw product.With preparation HPLC (Sunfire post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, water (containing 0.1%TFA) that reduces gradually with polarity and the mixture of MeCN are as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is solid (91mg, 50.4%); ¹H NMR (400.132MHz, DMSO) δ 1.53-1.61 (1H, m), 1.66-1.73 (1H, m), and 1.88-1.94 (1H, m), 2.02-2.09 (1H, m), and 3.14-3.25 (1H, m), 3.32-3.48 (2H, m), and 3.98-4.07 (1H, m), 4.34 (2H, s), and 5.33-5.39 (1H, m), 6.95 (1H, d), and 7.21-7.25 (1H, m), 7.37-7.44 (2H, m), and 7.80-7.84 (1H, m), 7.87-7.90 (1H, m), 7.97 (1H, d), 8.26 (1H, d), 8.54 (1H, d), 8.79 (1H, s), 12.56 (1H, s); M/z (LC-MS, ESI+), RT=1.69 (M+H 460.6).

(b) multiple parallel synthetic (26b-m)

According to above-mentioned steps, (150mg 0.39mmol) with suitable chlorine heterogeneous ring compound reaction, obtains required compound to make 4-(4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (2a)

26b：- ¹H?NMR(400.132MHz，DMSO)δ1.55-1.64(1H，m)，1.68-1.78(1H，m)，1.91-1.99(1H，m)，2.07-2.13(1H，m)，2.55(3H，s)，3.18-3.26(1H，m)，3.34-3.42(1H，m)，3.45-3.53(1H，m)，3.97-4.06(1H，m)，4.34(2H，s)，5.36-5.42(1H，m)，7.21-7.28(2H，m)，7.36-7.39(1H，m)，7.40-7.44(1H，m)，7.65(1H，d)，7.80-7.83(1H，m)，7.86-7.90(1H，m)，7.98(1H，d)，8.26(1H，d)，12.56(1H，s)

26c：- ¹H?NMR(400.132MHz，DMSO)δ1.51-1.62(1H，m)，1.64-1.73(1H，m)，1.87-1.93(1H，m)，2.02-2.09(1H，m)，3.17-3.24(1H，m)，3.32-3.49(2H，m)，3.99-4.06(1H，m)，4.34(2H，s)，5.31-5.37(1H，m)，7.00(1H，d)，7.20-7.25(1H，m)，7.36-7.38(1H，m)，7.40-7.44(1H，m)，7.80-7.84(1H，m)，7.86-7.90(1H，m)，7.97(1H，d)，8.16(1H，d)，8.26(1H，d)，8.69(1H，s)，12.56(1H，s)

26d：- ¹H?NMR(400.132MHz，CDCl3)δ1.79-2.16(4H，m)，3.29-3.37(1H，m)，3.52-3.63(1H，m)，3.74-3.87(1H，m)，3.97-4.07(1H，m)，4.33(2H，s)，5.32-5.39(1H，m)，7.04-7.08(1H，m)，7.29-7.33(1H，m)，7.35-7.37(1H，m)，7.79-7.87(3H，m)，8.14(1H，d)，8.18(1H，s)，8.24(1H，s)，8.48(1H，d)，11.30(1H，s)

26e：- ¹H?NMR(400.132MHz，DMSO)δ1.55-1.74(2H，m)，1.89-1.94(1H，m)，2.04-2.10(1H，m)，3.18-3.24(1H，m)，3.34-3.48(2H，m)，3.99-4.06(1H，m)，4.34(2H，s)，5.19-5.25(1H，m)，7.12-7.15(1H，m)，7.21-7.25(1H，m)，7.39-7.43(2H，m)，7.80-7.83(1H，m)，7.86-7.90(1H，m)，7.98(1H，d)，8.26(1H，d)，8.61(2H，d)，12.57(1H，s)

26f：- ¹H?NMR(400.132MHz，DMSO)δ1.51-1.59(1H，m)，1.64-1.73(1H，m)，1.86-1.93(1H，m)，2.02-2.08(1H，m)，3.17-3.22(1H，m)，3.34-3.47(2H，m)，3.98-4.07(1H，m)，4.34(2H，s)，5.26-5.32(1H，m)，7.21-7.25(1H，m)，7.36-7.44(4H，m)，7.80-7.84(1H，m)，7.86-7.90(1H，m)，7.97(1H，d)，8.25(1H，d)，8.40(1H，d)，12.57(1H，s)

26g：- ¹H?NMR(400.132MHz，DMSO)δ1.51-1.60(1H，m)，1.64-1.72(1H，m)，1.87-1.93(1H，m)，2.02-2.09(1H，m)，3.19-3.24(1H，m)，3.34-3.49(2H，m)，3.99-4.07(1H，m)，4.34(2H，s)，5.26-5.31(1H，m)，7.19(1H，s)，7.21-7.25(1H，m)，7.34-7.44(3H，m)，7.64(1H，s)，7.80-7.83(1H，m)，7.87-7.90(1H，m)，7.98(1H，d)，8.13(1H，s)，8.25-8.27(2H，m)，12.56(1H，s)

26h：- ¹H?NMR(400.132MHz，DMSO)δ1.57-1.63(1H，m)，1.71-1.77(1H，m)，1.83-1.90(1H，m)，2.00-2.06(1H，m)，2.40(3H，s)，3.23-3.28(1H，m)，3.34-3.41(1H，m)，3.59-3.64(1H，m)，3.85-3.91(1H，m)，4.34(2H，s)，5.27-5.31(1H，m)，7.21-7.25(1H，m)，7.36-7.38(1H，m)，7.41-7.44(1H，m)，7.79-7.83(1H，m)，7.86-7.90(1H，m)，7.96-8.01(2H，m)，8.05(1H，d)，8.25(1H，d)，12.56(1H，s)

26i：- ¹H?NMR(400.132MHz，CDCl3)δ1.61-1.67(1H，m)，1.72-1.80(1H，m)，1.88-1.95(1H，m)，2.03-2.10(1H，m)，2.42(3H，s)，2.44(3H，s)，3.25-3.32(1H，m)，3.47-3.55(1H，m)，3.76-3.99(2H，m)，4.29(2H，s)，5.26-5.31(1H，m)，7.01-7.05(1H，m)，7.27-7.31(1H，m)，7.33-7.35(1H，m)，7.71-7.79(4H，m)，8.46(1H，d)，10.45(1H，s)

26j：- ¹H?NMR(400.132MHz，DMSO)δ1.55-1.63(1H，m)，1.69-1.77(1H，m)，1.81-1.89(1H，m)，1.97-2.05(1H，m)，2.35(6H，s)，3.20-3.25(1H，m)，3.34-3.39(1H，m)，3.60-3.69(1H，m)，3.80-3.88(1H，m)，4.34(2H，s)，5.27-5.32(1H，m)，7.21-7.25(1H，m)，7.37(1H，d)，7.41-7.45(1H，m)，7.79-7.83(1H，m)，7.87-7.90(1H，m)，7.92(1H，s)，7.97(1H，d)，8.25(1H，d)，12.56(1H，s)

26k：- ¹H?NMR(400.132MHz，DMSO)δ1.46-1.56(1H，m)，1.60-1.68(1H，m)，1.82-1.91(1H，m)，1.98-2.06(1H，m)，3.13-3.22(1H，m)，3.32-3.45(2H，m)，3.80(3H，s)，3.99-4.07(1H，m)，4.34(2H，s)，5.22-5.26(1H，m)，6.33(1H，s)，6.57-6.59(1H，m)，7.20-7.24(1H，m)，7.36-7.44(2H，m)，7.80-7.84(1H，m)，7.86-7.91(1H，m)，7.94-7.99(2H，m)，8.26(1H，d)，12.56(1H，s)

26l：- ¹H?NMR(400.132MHz，DMSO)δ1.66-1.72(1H，m)，1.77-1.83(1H，m)，1.89-1.96(1H，m)，2.05-2.12(1H，m)，3.21-3.42(2H，m)，3.55-3.65(1H，m)，3.88-3.97(1H，m)，4.34(2H，s)，5.40-5.44(1H，m)，7.21-7.25(1H，m)，7.40-7.43(2H，m)，7.80-7.84(1H，m)，7.87-7.91(1H，m)，7.99(1H，d)，8.26(1H，d)，8.44(1H，d)，8.58(1H，d)，12.56(1H，s)

26m：- ¹H?NMR(400.132MHz，DMSO)δ1.48-1.55(1H，m)，1.60-1.66(1H，m)，1.81-1.88(1H，m)，1.98-2.03(1H，m)，3.11-3.20(1H，m)，3.33-3.48(2H，m)，3.93-4.02(1H，m)，4.33(2H，s)，4.70-4.76(1H，m)，7.20-7.25(1H，m)，7.36-7.45(3H，m)，7.54-7.57(1H，m)，7.80-7.84(1H，m)，7.87-7.90(1H，m)，7.97(1H，d)，8.16(1H，s)，8.26(1H，d)，12.57(1H，s)

Embodiment 17

(a) 4-(1,3-difluoro third-2-base oxygen base) pyridine (27)

Under 20 ℃, air atmosphere, with azo-2-carboxylic acid's di tert butyl carbonate (719mg, 3.12mmol) disposable join pyridine-4-alcohol (YY) (297mg, 3.12mmol), 1, (100mg is 1.04mmol) with polymkeric substance loading type triphenylphosphine (1.89mmol/g for 3-difluoro propan-2-ol; 1016mg is in methylene dichloride 3.12mmol) (10ml) solution.The suspension that obtains was stirred 4 hours down at 20 ℃.Mixture is filtered, and, obtain yellow oil solvent evaporation.With methylene dichloride (25ml) diluted reaction mixture, and use successively 2M NaOH (2 * 10ml) and saturated brine (10ml) clean.Organic layer passes through MgSO ₄Drying is filtered and evaporation, obtains raw product.Raw product is dissolved in methylene dichloride (25ml) again, and cleans with 2MHCl (10ml).With 2M NaOH alkalization water layer, and extract with methylene dichloride (* 3).The organic layer that merges passes through MgSO ₄Drying is filtered and evaporation, obtains required compound, is viscous oil (90mg, 50%); ¹H NMR (400.132MHz, CDCl ₃) δ 4.60-4.67 (2H, m), 4.71-4.76 (2H, m), 4.78-4.90 (1H, m), 6.92 (2H, d), 8.48 (2H, d); M/z (LC-MS, ESI+), RT=1.53 (M+H 174).

(b) 4-(1,3-difluoro third-2-base oxygen base) piperidines (28)

Under 25 ℃, the nitrogen atmosphere of 5bar, with MeOH (50ml) solution stirring of 4-(1,3-difluoro third-2-base oxygen base) pyridine (27) (0.098mg, 0.57 μ mol) and 5% rhodium aluminum oxide (0.02g, 2.56 μ mol) 16 hours.Catalyzer is filtered and clean,, obtain required compound, be gelatinoid (0.082g, 81%) solvent evaporation with MeOH; ¹H NMR (400.132MHz, CDCl ₃) δ 1.43-1.48 (1H, m), 1.60-1.71 (1H, m), 1.82-2.15 (3H, m), 2.42 (1H, s), and 2.84-2.92 (1H, m), 3.09-3.14 (1H, m), 3.25-3.30 (1H, m), 3.82-3.95 (1H, m), 4.35-4.44 (2H, m), 4.47-4.56 (2H, m).

(c) 4-(3-(4-(1,3-difluoro third-2-base oxygen base) piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (29)

Under 20 ℃, air atmosphere, with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (0.227g, 0.60mmol) disposable join 2-fluoro-5-((4-oxygen-3,4-dihydro-phthalazines-1-yl) methyl) phenylformic acid (1) (0.137g, 0.46mmol), 4-(1,3-difluoro third-2-base oxygen base) piperidines (28) (0.082g, 0.46mmol) and triethylamine (0.194ml, N 1.38mmol) is in dinethylformamide (3ml) solution.The solution that obtains was stirred 24 hours down at 20 ℃.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude mixture, the water that reduces gradually with polarity (contains 0.1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is colourless jelly (34mg, 15.90%); ¹H NMR (400.132MHz, DMSO) δ 1.10-1.19 (1H, m), 1.21-1.30 (1H, m), 1.46-1.55 (2H, m), and 1.62-1.70 (2H, m), 2.78-2.86 (1H, m), 3.54-3.60 (1H, m), 3.69-3.79 (2H, m), 4.11 (2H, s), 4.15-4.20 (1H, m), and 4.22-4.32 (2H, m), 4.34-4.39 (1H, m), 6.97-7.02 (1H, m), and 7.13-7.20 (2H, m), 7.59-7.69 (2H, m), 7.76 (1H, d), 8.05 (1H, d), 12.34 (1H, s); M/z (LC-MS, ESI+), RT=1.99 (M+H 460).

Embodiment 18

(a) (Z)-3-(3-bromo-4 fluorobenzene methylene radical)-7-nitro isobenzofuran-1 (3H)-ketone (30a) and (Z)-3-(3-bromo-4-fluorobenzene methylene radical)-4-nitro isobenzofuran-1 (3H)-ketone (30b)

Under air atmosphere, with sodium acetate (0.018g 0.21mmol) joins 4-nitro isobenzofuran-1, the 3-diketone (1.409g, 7.30mmol) and 2-(3-bromo-4-fluorophenyl) acetate (1g, 4.29mmol) in.The mixture that obtains was stirred 30 minutes at 240 ℃.The refrigerative mixture is pulverized with ethanol, filtered and drying, obtain the mixture of required compound, be yellow solid (0.111g, 3.55%); M/z (LC-MS, ESI+), RT=2.92 (78.7%) and 3.02 (21.3%).It need not to be further purified when using in next stage.

To evaporate by the ethanolic soln that filtration obtains, obtain dark-brown remaining jelly, use the silicon-dioxide chromatogram purification, use 0-100%EtOAc/ isohexane wash-out, obtain (Z)-3-(3-bromo-4-fluorobenzene methylene radical)-4-nitro isobenzofuran-1 (3H)-ketone (JME3a ') (45mg, 2.9%); ¹H NMR (400.132MHz, DMSO) δ 7.16 (1H, s), 7.51-7.56 (1H, m), 7.84-7.89 (1H, m), 8.10-8.14 (2H, m), 8.21 (1H, d), 8.40 (1H, d); M/z (LC-MS, ESI+), RT=2.97 (not finding M+H); (Z)-3-(3-bromo-4-fluorobenzene methylene radical)-7-nitro isobenzofuran-1 (3H)-ketone (JME3a) (4mg, 0.3%); ¹H NMR (400.132MHz, DMSO) δ 7.12 (1H, s), 7.50-7.54 (1H, m), 7.90-7.95 (2H, m), 8.21-8.23 (1H, m), 8.37 (1H, d), 8.53 (1H, d); M/z (LC-MS, ESI+), RT=3.07 (not finding M+H).

(b) 4-(3-bromo-4-luorobenzyl)-5-nitro phthalazines-1 (2H)-ketone compound (31b) and 4-(3-bromo-4-luorobenzyl)-8-nitro phthalazines-1 (2H)-ketone (31a)

Under 20 ℃, air atmosphere, with hydrazine hydrate (0.215ml, 4.43mmol) join (Z)-3-(3-bromo-4 fluorobenzene methylene radical)-4-nitro isobenzofuran-1 (3H)-ketone (30b) with (Z)-mixture (403mg of 3-(3-bromo-4-fluorobenzene methylene radical)-7-nitro isobenzofuran-1 (3H)-ketone (30a) (4: 1), 0.55mmol) and N, (0.214ml is in water 2.77mmol) (10ml) solution for dinethylformamide.The mixture that obtains was stirred 18 hours down at 100 ℃.With solvent evaporation, and resistates is dissolved in methylene dichloride/MeOH.Reduce solvent volume, obtain the mixture of 4-(3-bromo-4-luorobenzyl)-5-nitro phthalazines-1 (2H)-ketone and 4-(3-bromo-4-luorobenzyl)-8-nitro phthalazines-1 (2H)-ketone,, its taking-up be need not to be further purified for dark brown solid (429mg, 103%); ¹H NMR (400.132MHz, DMSO) δ 4.37 ﹠amp; (4.05 2H, 2 x s), 7.20-7.39 (2H, m), 7.53-7.72 (1H, m), 7.92-8.40 (3H, m), 9.04 (1H, s); M/z (LC-MS, ESI+), RT=2.38 (M+H 380).

(c) 2-fluoro-5-((5-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) cyanobenzene (32b) and 2-fluoro-5-((8-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) cyanobenzene (32a)

Under 20 ℃, air atmosphere, with cuprous cyanide (I) (71.7mg, 0.80mmol) join 4-(3-bromo-4-luorobenzyl)-5-nitro phthalazines-1 (2H)-ketone (31b) and 4-(3-bromo-4-luorobenzyl)-8-nitro phthalazines-1 (2H)-ketone (31a) mixture (429mg, 0.57mmol) N, in dinethylformamide (4ml) solution.The brown solution that obtains was stirred 17 hours down at 160 ℃.(11mg 0.12mmol) also continues reaction 5 hours to add cuprous cyanide (I).The refrigerative mixture is diluted with ethyl acetate (50ml), and (2 * 25ml) clean with saturated brine.Pass through MgSO ₄Dry organic solution is filtered and evaporation, obtains brown jelly (150mg, productive rate 40.8%), and it need not to be further purified in use; M/z (LC-MS, ESI+), RT=2.05 (M-H 323).

(d) 2-fluoro-5-((5-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (33b) and 2-fluoro-5-((8-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (33a)

Under 20 ℃, air atmosphere, with potassium hydroxide (14mg, 0.25mmol) water (1ml) solution join 2-fluoro-5-((5-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) cyanobenzene (32a) and 2-fluoro-5-((8-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) (162mg is in ethanol 0.25mmol) (4ml) solution for cyanobenzene (32b) mixture.The solution that obtains was at room temperature stirred 2 hours, and stirred 2 hours down at 50 ℃.And then (93mg, 1.66mmol 7eq) and at 90 ℃ heated 16 hours down to add potassium hydroxide.With the reaction mixture cooling, and water (10ml) dilution, with diethyl ether (2 * 10ml) extractions.With the 1M citric acid acidifying aqueous solution, solid product is filtered, 60 ℃ of following vacuum-dryings, obtain desired mixt (26mg, productive rate 15%).Extract remaining aqueous solution with EtOAc, the extraction liquid that merges is passed through dried over mgso, filter also evaporation, obtain more desired mixt, be jelly (27mg, productive rate 15%); M/z (LC-MS, ESI+), RT=0.84 (M+H 344).Two batches of products are merged, and need not in use to be further purified.

(e) 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5-nitro phthalazines-1 (2H)-ketone (34b) and 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-8-nitro phthalazines-1 (2H)-ketone (34a)

Under 20 ℃, air atmosphere, with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (38.1mg, 0.10mmol) disposable 2-fluoro-5-((the 5-nitro-4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) methyl) phenylformic acid (33b) and 2-fluoro-5-((8-nitro-4-oxygen-3,4-dihydro phthalazines-1-yl) mixture (53mg of phenylformic acid (33a) methyl), 0.08mmol), 4-methoxyl group piperidines (17.8mg, 0.15mmol) and triethylamine (0.033ml is in DMF 0.24mmol) (2ml) solution.The solution that obtains was stirred 24 hours down at 20 ℃.Use the ethyl acetate diluted reaction mixture, and water (* 2) cleans.Pass through MgSO ₄Dry organic layer filters and evaporation, obtains desired mixt, is jelly (8.5mg, productive rate 12.50%) that it need not to be further purified in use; M/z (LC-MS, ESI+), RT=1.76 (M+H 441.3).

(f) 8-amino-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (35a)

Under 20 ℃, with 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5-nitro phthalazines-1 (2H)-ketone (34b) and 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-8-nitro phthalazines-1 (2H)-ketone (34a) mixture (6mg, 6.81 ethanol μ mol) (2ml) solution joins 5% palladium-carbon catalyst (2mg, 0.94 in ethanol μ mol) (5ml) solution, and the mixture that obtains stirred 5 hours under 20 ℃, nitrogen atmosphere.Catalyzer is filtered and clean with ethanol.Evaporating solvent obtains foam.Add ethanol and 5% palladium carbon (10mg) again, and with mixture restir 5 hours under nitrogen atmosphere.Catalyzer is filtered, and clean with ethanol.Evaporation obtains jelly, and with preparation HPLC (Waters XBridge Prep C18OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) this jelly of purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is colourless jelly (1.700mg, 60.8%); ¹H NMR (400.132MHz, CDCl3) δ 1.25-1.31 (1H, m), 1.51-1.82 (2H, m), 1.89-1.98 (1H, m), 3.09-3.17 (1H, m), 3.38 (3H, s), 3.46-3.51 (2H, m), and 3.55-3.63 (1H, m), 3.98-4.05 (1H, m), 4.16 (2H, s), 6.43 (2H, s), 6.79 (1H, d), 6.85 (1H, d), 7.00-7.04 (1H, m), 7.25-7.31 (2H, m), 7.41-7.45 (1H, m), 9.56 (1H, s); M/z (LC-MS, ESI+), RT=1.71 (M+H 411.5).

Embodiment 19

4-(3-(4-(cyclopentyloxy) piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (36)

Under 20 ℃, air atmosphere, with triethylamine (0.280ml, 2.01mmol) disposable 2-fluoro-5-((the 4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (200mg, 0.67mmol), 4-(cyclopentyloxy) piperidine hydrochlorate (113mg, 0.67mmol) and O-benzotriazole-1-base-N, N, N ', (331mg is in DMA 0.87mmol) (2ml) solution for N '-tetramethyl-urea hexafluorophosphate.The mixture that obtains was stirred 3 days down at 20 ℃.With preparation HPLC (Waters XBridge Prep C18OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, water (containing 1%NH3) that reduces gradually with polarity and the mixture of MeCN are as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (112mg, productive rate 37.2%); ¹H NMR (400.132MHz, DMSO) δ 1.23-1.72 (11H, m), 1.81-1.87 (1H, m), 3.00-3.06 (1H, m), 3.22-3.27 (2H, m), 3.52-3.59 (1H, m), 3.96-4.04 (2H, m), 4.33 (2H, s), 7.18-7.23 (1H, m), 7.34 (1H, d), 7.38-7.42 (1H, m), 7.81-7.91 (2H, m), 7.97 (1H, d), 8.26 (1H, d), 12.56 (1H, s); M/z (LC-MS, ESI+), RT=2.30 (M+H 450.1).

Embodiment 20

4-(3-(4-(cyclopentyloxy) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (37)

Under 20 ℃, air atmosphere, with triethylamine (0.298ml, 2.14mmol) disposable 3-((the 4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1b) (200mg, 0.71mmol), 4-(cyclopentyloxy) piperidine hydrochlorate (121mg, 0.71mmol) and O-benzotriazole-1-base-N, N, N ', (352mg is in DMA 0.93mmol) (2ml) solution for N '-tetramethyl-urea hexafluorophosphate.The mixture that obtains was stirred 3 days down at 20 ℃.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (120mg, productive rate 39%); ¹H NMR (400.132MHz, DMSO) δ 1.22-1.86 (11H, m), 3.00-3.20 (2H, m), 3.25-3.43 (2H, m), 3.52-3.56 (1H, m), 3.91-4.05 (2H, m), 4.35 (2H, s), 7.20 (1H, d), 7.31 (1H, s), 7.34-7.40 (2H, m), 7.80-7.89 (2H, m), 7.96 (1H, d), 8.26 (1H, d), 12.57 (1H, s); M/z (LC-MS, ESI+), RT=2.18 (M+H 432.1)

Embodiment 21

4-(4-fluoro-3-(4-(6-picoline-2-base oxygen base) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (39)

With 6-picoline-2 alcohol (95mg, 0.87mmol) the disposable polymkeric substance loading type triphenylphosphine (1.89mmol/g that joins; 460mg is in DCM 0.87mmol) (5ml) solution.Add azo-2-carboxylic acid's di tert butyl carbonate (201mg, 0.87mmol) and 4-(4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (2a) (111mg, 0.29mmol) DCM (1ml) solution, and the mixture that obtains stirred 4 hours down at 20 ℃.Use the DCM diluted reaction mixture, and water (* 2) cleans.Pass through MgSO ₄Dry organic layer filters and evaporation, obtains raw product.With preparation HPLC (WatersXBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, water (containing 1% formic acid) that reduces gradually with polarity and the mixture of MeCN are as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (10mg, productive rate 7.3%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.70-1.80 (1H, m), 1.85-1.95 (2H, m), 2.02-2.08 (1H, m), 2.41 (3H, s), and 3.22-3.28 (1H, m), 3.51-3.57 (1H, m), 3.74-3.83 (1H, m), 3.95-4.03 (1H, m), 4.28 (2H, s), 5.32-5.35 (1H, m), 6.50 (1H, d), 6.69 (1H, d), 7.02 (1H, t), 7.26-7.29 (1H, m), 7.33-7.35 (1H, m), 7.44 (1H, t), 7.71-7.79 (3H, m), 8.45-8.48 (1H, m), 10.33 (1H, s); M/z (LC-MS, ESI+), RT=2.26 (M+H 473.6).

Embodiment 22

4-((2-(4-(cyclopentyloxy) piperidines-1-carbonyl) pyridin-4-yl) methyl) phthalazines-1 (2H)-ketone (40)

Under 20 ℃, air atmosphere, with triethylamine (0.452ml, 3.24mmol) disposable 4-((the 4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) methyl) pyridine carboxylic acid (5) (304mg, 1.08mmol), 4-(cyclopentyloxy) piperidine hydrochlorate (183mg, 1.08mmol) and O-benzotriazole-1-base-N, N, N ', (533mg is in DMA 1.41mmol) (3ml) solution for N '-tetramethyl-urea hexafluorophosphate.The mixture that obtains was stirred 3 days down at 20 ℃.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (39mg, productive rate 8.3%); ¹H NMR (400.132MHz, DMSO) δ 1.28-1.74 (11H, m), 1.82-1.88 (1H, m), 3.08-3.14 (1H, m), 3.20-3.27 (1H, m), 3.44-3.49 (1H, m), and 3.53-3.59 (1H, m), 3.96-4.05 (2H, m), 4.40 (2H, s), 7.39 (1H, d), 7.49 (1H, s), 7.83-7.96 (3H, m), 8.28 (1H, d), 8.47 (1H, d), 12.60 (1H, s); M/z (LC-MS, ESI+), RT=1.88 (M+H 433.1).

Embodiment 23

(a) (2-fluoro-5-iodophenyl) (4-methoxyl group piperidines-1-yl) ketone (41)

With 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea hexafluorophosphate (V) (5.99g, 15.79mmol) join 2-fluoro-5-iodo-benzoic acid (3.000g in batches, 11.28mmol), 4-methoxyl group piperidines (1.364g, 11.84mmol) and triethylamine (3.93ml is in DMF 28.19mmol) (40ml) solution.The solution that obtains was at room temperature stirred 7 hours.Reaction mixture is poured in the water (350ml), and (2 * 200ml) extractions are cleaned the organic layer that merges with saturated brine (100ml), use Na with Et2O ₂SO ₄Drying is filtered and evaporation, obtains the darkorange jelly.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-50%EtOAc.Pure fraction is evaporated to drying, obtains desired substance, be colourless jelly (0.348g, productive rate 67%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.57-1.99 (4H, m), 3.18 (1H, m), 3.39 (3H, s), 3.50 (2H, m), 3.63 (1H, m), 3.98 (1H, m), 6.89 (1H, t), 7.69 (2H, m); M/z (LC-MS, ESI+), RT=2.05 (M+H364.0).

(b) 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethyl acetate (42)

Under 25 ℃, nitrogen atmosphere, with cesium carbonate (2.308g, 7.08mmol) disposable joining (2-fluoro-5-iodophenyl) (4-methoxyl group piperidines-1-yl) ketone (41) (1.06g, 2.92mmol), diethyl malonate (1.329ml, 8.76mmol), 2-phenylphenol (0.099g, 0.58mmol) and cuprous iodide (I) (0.056g is in THF 0.29mmol) (10ml) solution.The mixture that obtains was stirred 90 minutes in microwave reactor under 160 ℃.Reaction mixture is evaporated to drying, and is dissolved in Et again ₂O (200ml), water (100ml) and saturated brine (50ml) clean successively.Pass through MgSO ₄Dry organic layer filters and evaporation, obtains raw product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-50% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be colourless gelatinoid (2.03g, productive rate 72%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.25 (3H, t), 1.53-1.61 (1H, m), 1.67-1.74 (1H, m), and 1.77-1.84 (1H, m), 1.89-1.97 (1H, m), 3.12-3.20 (1H, m), 3.36 (3H, s), 3.44-3.62 (3H, m), 3.59 (2H, s), 3.96-4.05 (1H, m), 4.15 (2H, q), 7.02-7.06 (1H, m), 7.26-7.33 (2H, m); M/z (LC-MS, ESI+), RT=1.08 (M+H 324.3).

(c) 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) acetate (43)

Under 25 ℃, air atmosphere, (0.752g 31.39mmol) disposablely joins 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethyl acetate (42) (2.03g is in THF 6.28mmol) (20ml) and water (20ml) solution with lithium hydroxide.The mixture that obtains was at room temperature stirred 2 hours.With the THF evaporation, and water (30ml) diluted mixture thing.With 2N HCl acidified aqueous solution, and with EtOAc (3 * 30ml) extraction products.Dry (MgSO ₄) extract that merges, filter also evaporation, obtain desired substance, be white solid (1.580g, productive rate 85%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.54-1.84 (3H, m), 1.89-1.96 (1H, m), 3.12-3.20 (1H, m), 3.36 (3H, s), 3.45-3.53 (2H, m), 3.58 (2H, s), 3.60-3.66 (1H, m), 3.93-4.02 (1H, m), 7.01-7.06 (1H, m), 7.28-7.32 (2H, m); M/z (LC-MS, ESI+), RT=0.78 (M+H 296.3).

(d) (Z)-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji)-4-nitro isobenzofuran-1 (3H)-ketone (44)

Under air atmosphere, with sodium acetate (6.97mg 0.08mmol) joins 4-nitro isobenzofuran-1, the 3-diketone (558mg, 2.89mmol) and 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) acetate (43) (502mg, 1.70mmol) in.The mixture that obtains was stirred 30 minutes down at 240 ℃.With quick silicon-dioxide chromatogram purification refrigerative mixture, gradient is the isohexane solution of 0-100% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be yellow jelly matter (50mg, productive rate 6.90%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.59-2.00 (4H, m), 3.10-3.26 (1H, m), 3.37 (3H, s), and 3.43-3.57 (2H, m), 3.61-3.71 (1H, m), 3.88-4.05 (1H, m), 7.16-7.20 (1H, m), 7.30 (1H, s), 7.70-7.76 (1H, m), 7.86-7.88 (1H, m), 7.97-8.01 (1H, m), 8.27 (1H, d), 8.39 (1H, d); M/z (LC-MS, ESI+), RT=2.29 (M+H 427.3).

(e) 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5-nitro phthalazines-1 (2H)-ketone (45)

Under 25 ℃, air atmosphere, with hydrazine hydrate (0.046ml, 0.94mmol) disposable joining (Z)-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji)-4-nitro isobenzofuran-1 (3H)-ketone (44) (50mg, 0.12mmol) and N, (0.045ml is in water 0.59mmol) (2ml) solution for dinethylformamide.The mixture that obtains was stirred 18 hours down at 100 ℃.Mixture is cooled to room temperature, and, obtains rough required product, be light brown solid (40.0mg, productive rate 77%) solvent evaporation; ¹H NMR:(400.132MHz, DMSO) δ 1.23-1.33 (1H, m), 1.36-1.50 (1H, m), 1.65-1.72 (1H, m), 1.79-1.92 (1H, m), 3.25-3.27 (4H, m), 3.33 (3H, s), 3.82-3.97 (1H, m), 4.10 (2H, s), 7.10-7.21 (3H, m), 7.99-8.03 (1H, m), 8.27 (1H, d), 8.59 (1H, d); M/z (LC-MS, ESI+), RT=1.85 (M+H 441.3).

(f) 5-amino-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1-(2H)-ketone (46)

Under 25 ℃, air atmosphere, (40mg, (4mg is in ethanol 0.04mmol) (7ml) solution 0.09mmol) to join 5% palladium carbon with 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5-nitro phthalazines-1 (2H)-ketone (45).With the mixture that obtains 25 ℃ of following hydrogenations 22 hours.Catalyzer is filtered and clean,, obtain the light brown jelly solvent evaporation with ethanol.With preparation HPLC (WatersXBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is light brown solid (11mg, productive rate 29.5%); ¹H NMR (400.132MHz, DMSO) δ 1.66-1.81 (2H, m), 1.86-1.96 (2H, m), 3.10-3.19 (1H, m), 3.36 (3H, s), 3.44-3.50 (2H, m), 3.55-3.64 (1H, m), 3.91-3.99 (1H, m), 4.08 (2H, s), 4.44 (2H, s), 6.99 (1H, d), 7.05-7.10 (1H, m), 7.23-7.28 (2H, m), 7.50-7.54 (1H, m), 7.94 (1H, d), 9.95 (1H, s); M/z (LC-MS, ESI+), RT=1.50 (M+H 411.3).

Embodiment 24

(a) 4-(3-(4-oxyethyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (47a)

With 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea hexafluorophosphate (V) (0.379g, 1.00mmol) join 3-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1b) (0.200g, 0.71mmol), 4-oxyethyl group piperidines (0.101g, 0.78mmol) and triethylamine (0.249ml is in DMF 1.78mmol) (3ml) solution.The solution that obtains was at room temperature stirred 3 days.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound.(0.175g, productive rate 62.6%); ¹H NMR (400.132MHz, DMSO) δ 1.11 (3H, t), 1.20-1.93 (4H, m), 2.98-3.27 (2H, m), 3.33-3.41 (1H, m), 3.42-3.54 (3H, m), 3.93 (1H, s), 4.35 (2H, s), 7.21 (1H, m), 7.31 (1H, s), 7.33-7.42 (2H, m), 7.79-7.90 (2H, m), 7.95 (1H, m), 8.26 (1H, m), 12.58 (1H, s); M/z (LC-MS, ESI+), RT=1.81 minute (M+H=392.13).

(b) 4-(3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (47b)

With 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea hexafluorophosphate (V) (0.379g, 1.00mmol) join 3-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1b) (0.200g, 0.71mmol), 4-methoxyl group piperidines (0.090g, 0.78mmol) and triethylamine (0.249ml is in DMF 1.78mmol) (3ml) solution.The solution that obtains was at room temperature stirred 16 hours.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (0.211g, productive rate 78%); ¹H NMR (400.132MHz, DMSO) δ 1.24-1.49 (2H, m), 1.63-1.93 (2H, m), 2.94-3.14 (2H, m), 3.30 (3H, s), 3.34-3.44 (2H, m), 3.88 (1H, s), 4.35 (2H, s), 7.21 (1H, m), 7.30-7.43 (3H, m), 7.80-7.90 (2H, m), 7.96 (1H, d), 8.27 (1H, m), 12.58 (1H, s); M/z (LC-MS, ESI+), RT=1.57 minute (M+H=378.20).

Embodiment 25

(a) 2-(3-bromophenyl) Acetyl Chloride 98Min. (48)

(2.035ml, (1.500g is in DCM 6.98mmol) (30ml) solution 27.90mmol) to join 2-(3-bromophenyl) acetate with thionyl chloride.With to solution at room temperature stirred 18 hours.The mixture that obtains is evaporated to drying, and with resistates and methylbenzene azeotropic, obtains raw product (1.570g, productive rate 96%), it need not to be further purified in use.

(b) 4-(2-(3-bromophenyl) ethanoyl)-2,5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (49)

Under 0 ℃, nitrogen atmosphere, with 2, (0.295g, anhydrous DCM (5ml) solution 1.93mmol) dropwise join the aluminum chloride that is stirring, and (0.714g is in anhydrous DCM (10ml) suspension 5.35mmol) for 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester.The suspension that obtains was stirred 10 minutes down at 0 ℃.Under 0 ℃, in the described suspension that is stirring, dropwise add 2-(3-bromophenyl) Acetyl Chloride 98Min. (48) (0.500g, anhydrous DCM (5ml) solution 2.14mmol).The suspension that obtains is warming up to room temperature, and at room temperature further stirs 18 hours.Reaction mixture is poured among ice (50ml) and the dense HCl (2ml).Separate each layer, and with DCM (2 * 50ml) aqueous layer extracted.Use successively 2M HCl (2 * 75ml), water (50ml) and saturated NaHCO ₃(50ml) clean the organism that merges.With organic layer Na ₂SO ₄Drying is filtered and evaporation, obtains raw product, and it directly uses in next stage; M/z (LC-MS, ESI-), RT=2.46 minute (M-H=350.00).

(c) 4-(3-bromobenzyl)-5,7-dimethyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (50)

(0.110ml 1.48mmol) joins 4-(2-(3-bromophenyl) ethanoyl)-2, and (0.470g is in acetate 1.34mmol) (20ml) solution for 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (49) with hydrazine hydrate.The solution that obtains was at room temperature stirred 2 days.Reaction mixture is evaporated to drying also is dissolved in again among the EtOAc (200ml), use 2M NaOH (100ml) and saturated brine (50ml) to clean successively.Organic layer is passed through Na ₂SO ₄Drying is filtered and evaporation, obtains raw product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be light brown solid (0.253g, productive rate 56.7%); ¹H NMR (400.132MHz, MeOD) δ 2.18 (3H, s), 2.50 (3H, s), 4.03 (2H, s), 7.04-7.11 (2H, m), 7.25 (2H, m); M/z (LC-MS, ESI+), RT=1.84 minute (M+H=333.97).

(d) 3-((5,7-dimethyl-1-oxygen-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-4-yl) methyl) phenylformic acid (51)

With 4-(3-bromobenzyl)-5,7-dimethyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (50) (0.188g, 0.57mmol), hexacarbonylmolybdenum (0.224g, 0.85mmol), N, N-lutidine-4 amine (0.138g, 1.13mmol), N-ethyl-N-sec.-propyl third-2-amine (0.197ml, 1.13mmol) and acetoxyl group (2-(di-o-tolyl phosphine) benzyl) palladium (0.027g, 0.03mmol) be suspended in the mixture of diox (2.0ml) and water (2.0ml), and be sealed in the microwave tube.In microwave reactor, will be reflected at 150 ℃ of heating 30 minutes, and be cooled to room temperature.Water (50ml) and EtOAc (50ml) diluted reaction mixture.By the diatomite filtration reaction mixture.Separate each layer, and the aqueous solution is adjusted to pH2 with 2M HCl.Collect the precipitation that obtains, water (50ml), Et by filtering ₂O (50ml) cleans, and dry under vacuum, obtains desired substance, is brown solid (0.200g, productive rate＞100%) that it need not to be further purified when using. ¹H NMR (400.132MHz, DMSO) δ 2.26 (3H, s), 2.51 (3H, s), 4.13 (2H, s), 7.39-7.48 (2H, m), 7.78 (2H, m), 11.08 (1H, s), 11.93 (1H, s), 12.86 (1H, s); M/z (LC-MS, ESI+), RT=1.41 minute (M+H=298.08).

(e) 4-(3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5,7-dimethyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (52a)

With 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.179g 0.47mmol) joins 3-((5 to 3-tetramethyl-isourea hexafluorophosphate (V), 7-dimethyl-1-oxygen-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-4-yl) methyl) phenylformic acid (51) (0.100g, 0.34mmol), 4-methoxyl group piperidines (0.043g, 0.37mmol) and triethylamine (0.117ml is in DMF 0.84mmol) (2ml) solution.The solution that obtains was at room temperature stirred 5 hours.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (0.054g, productive rate 40.9%). ¹H NMR (400.132MHz, DMSO) δ 1.18-1.93 (4H, m), 2.24 (3H, s), 2.51 (3H, s), 3.02-3.19 (2H, m), 3.25 (3H, s), 3.40 (2H, m), 3.76-3.97 (1H, m), 4.09 (2H, s), 7.14 (1H, s), 7.20 (1H, d), 7.27 (1H, d), 7.36 (1H, t), 11.08 (1H, s), 11.92 (1H, s); M/z (LC-MS, ESI+), RT=1.38 minute (M+H=395.09).

(f) 4-(3-(4-oxyethyl group piperidines-1-carbonyl) benzyl)-5,7-dimethyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (52b)

With 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.163g 0.43mmol) joins 3-((5 to 3-tetramethyl-isourea hexafluorophosphate (V), 7-dimethyl-1-oxygen-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-4-yl) methyl) phenylformic acid (51) (0.091g, 0.31mmol), 4-oxyethyl group piperidines (0.044g, 0.34mmol) and triethylamine (0.107ml is in DMF 0.77mmol) (2ml) solution.The solution that obtains was at room temperature stirred 16 hours.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance (0.028g, productive rate 22.5%). ¹H NMR (400.132MHz, DMSO) δ 1.11 (3H, t), 1.16-1.95 (4H, m), 2.24 (3H, s), 2.51 (3H, s), 2.99-3.26 (2H, m), and 3.33-3.54 (4H, m), 3.85-4.01 (1H, m), 4.09 (2H, s), 7.14 (1H, s), 7.20 (1H, d), 7.27 (1H, d), 7.35 (1H, t), 11.08 (1H, s), 11.92 (1H, s); M/z (LC-MS, ESI+), RT=1.53 minute (M+H=409.13).

Embodiment 26

(a) 4-(2-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-fluorophenyl) ethanoyl) 2,5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (53)

Under 0 ℃, nitrogen atmosphere, with 2, (0.545g, anhydrous DCM (10ml) solution 3.56mmol) dropwise join the aluminum chloride that is stirring, and (1.078g is in anhydrous DCM (20ml) suspension 8.08mmol) for 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester.The suspension that obtains was stirred 10 minutes down at 0 ℃.In the described suspension that is stirring, dropwise add under 0 ℃ essential acyl chloride solution [before the preparation; Pass through down to stir at 0 ℃ with 1-chloro-N, N, (0.428ml 3.23mmol) joins 2-(3-(4-oxyethyl group piperidines-1-carbonyl) 4-fluorophenyl) acetate (1.000g to 2-trimethylammonium third-1-alkene-1-amine, 3.23mmol) anhydrous DCM (10ml) solution in, and stirred 2 hours].The suspension that obtains is warming up to room temperature also at room temperature further to be stirred 18 hours.Unreacted is complete, and (1.078g 8.08mmol) and with solution at room temperature further stirred 1 hour to add aluminum chloride again.Reaction mixture is poured among ice (100ml) and the dense HCl (5ml).Separate each layer, and with DCM (2 * 100ml) aqueous layer extracted.Use respectively 2M HCl (2 * 150ml), water (100ml) and saturated NaHCO ₃(100ml) clean the organism that merges.Organic layer is passed through Na ₂SO ₄Drying is filtered and evaporation, obtains rough product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be light brown solid (1.241g, productive rate 86%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.14 (3H, t), 1.42-1.66 (2H, m), 1.71 (1H, m), 1.85 (1H, m), 2.00 (3H, s), 2.32 (3H, s), 3.06 (1H, m), 3.39-3.52 (5H, m), 3.75 (3H, s), 3.93-4.02 (3H, m), 6.92 (1H, t), 7.07 (1H, m), 7.17 (1H, m), 8.43 (1H, s); M/z (LC-MS, ESI+), RT=2.02 minute (M+H=445.24).

(b) 4-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-luorobenzyl)-5,7-dimethyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (54)

(0.037ml 0.49mmol) joins 4-(2-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-fluorophenyl) ethanoyl)-2, and (0.200g is in acetate 0.45mmol) (8ml) solution for 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (53) with hydrazine hydrate.The solution that obtains was at room temperature stirred 2 days.The mixture that obtains is evaporated to drying, and with resistates and methylbenzene azeotropic, obtain rough jelly, described jelly is pulverized with the mixture of NMP, DMSO and water, obtained solid, collect described solid by filtering, clean and drying under vacuum with ether, obtain required compound, be white solid (0.112g, productive rate 58.4%); ¹H NMR (400.132MHz, DMSO) δ 1.11 (3H, t), 1.29 (1H, m), 1.42 (1H, m), 1.71 (1H, m), 1.86 (1H, m), 2.27 (3H, s), 2.51 (3H, s), 3.04 (1H, m), 3.24-3.39 (2H, m), 3.46 (2H, m), 3.52 (1H, m), 3.97 (1H, s), 4.07 (2H, s), 7.13-7.30 (3H, m), 11.07 (1H, s), 11.94 (1H, s); M/z (LC-MS, ESI+), RT=1.62 minute (M+H=427.12).

(c) 4-(2-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-fluorophenyl) ethanoyl)-1,2,5-trimethylammonium-1H-pyrroles-3-carboxylate methyl ester (55)

With methyl iodide (0.056ml; 0.90mmol) join 4-(2-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-fluorophenyl) ethanoyl) 2; 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (53) (0.200g; 0.45mmol) and salt of wormwood (0.249g is in DMF 1.80mmol) (10ml) solution.The suspension that obtains was at room temperature stirred 2 hours.Reaction mixture is poured in the water (75ml) also with EtOAc (3 * 50ml) extractions.Water (50ml) and saturated brine (50ml) clean the organic layer that merges.With organic layer Na ₂SO ₄Drying is filtered and evaporation, obtains rough product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be yellow jelly (0.158g, productive rate 77%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.14 (3H, t), 1.43-1.65 (2H, m), 1.72 (1H, m), 1.85 (1H, m), 2.05 (3H, s), 2.38 (3H, s), 3.06 (1H, m), 3.32 (3H, s), 3.45 (5H, m), 3.74 (3H, s), 3.91 (2H, s), 3.99 (1H, m), 6.93 (1H, t), 7.08 (1H, m), 7.17 (1H, m); M/z (LC-MS, ESI+), RT=2.15 minute (M+H=459.36).

(d) 4-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-luorobenzyl)-5,6,7-trimethylammonium-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (56)

(0.028ml 0.38mmol) joins 4-(2-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-fluorophenyl) ethanoyl)-1,2, and (0.158g is in acetate 0.34mmol) (4ml) solution for 5-trimethylammonium-1H-pyrroles-3-carboxylate methyl ester (55) with hydrazine hydrate.The solution that obtains was at room temperature stirred 2 days.The mixture that obtains is evaporated to drying, and, obtains raw product resistates and methylbenzene azeotropic, with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (0.049g, productive rate 32.3%); ¹H NMR (400.132MHz, DMSO) δ 1.11 (3H, t), 1.29 (1H, m), 1.41 (1H, m), 1.72 (1H, m), 1.86 (1H, m), 2.29 (3H, s), 2.60 (3H, s), 3.05 (1H, m), 3.28 (3H, s), 3.46 (2H, m), 3.53 (3H, s), 3.96 (1H, m), 4.12 (2H, s), 7.16-7.28 (3H, m), 11.17 (1H, s); M/z (LC-MS, ESI+), RT=1.78 minute (M+H=441.11).

Embodiment 27

(a) 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-2,5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (57)

Under 0 ℃, nitrogen atmosphere, with 2, (0.114g, anhydrous DCM (5ml) solution 0.74mmol) dropwise join the aluminum chloride that is stirring, and (0.226g is in anhydrous DCM (10ml) suspension 1.69mmol) for 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester.The suspension that obtains was stirred 10 minutes down at 0 ℃.In the described suspension that is stirring, dropwise add under 0 ℃ essential acyl chloride solution [before the preparation; Under 0 ℃ with 1-chloro-N, N, (0.090ml 0.68mmol) joins 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) acetate (43) (0.200g to 2-trimethylammonium third-1-alkene-1-amine, 0.68mmol) anhydrous DCM (5ml) solution in, and stirred 2 hours].The suspension that obtains is warming up to room temperature, at room temperature further stirred 18 hours then.Unreacted is complete, and (0.226g 1.69mmol) and with solution at room temperature further stirred 1 hour to add aluminum chloride again.Reaction mixture is poured among ice (50ml) and the dense HCl (2ml).Separate each layer, and with DCM (2 * 50ml) aqueous layer extracted.Use successively 2M HCl (2 * 75ml), water (50ml) and saturated NaHCO ₃(50ml) clean the organism that merges.Organic layer is passed through Na ₂SO ₄Drying is filtered and evaporation, obtains rough product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100%EtOAc.Pure fraction is evaporated to drying, obtains 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-2,5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (0.223g, productive rate 76%) is Off-white solid. ¹H NMR (400.132MHz, CDCl ₃) δ 1.57-1.90 (4H, m), 1.98 (3H, s), 2.31 (3H, s), 3.07 (1H, m), 3.29 (3H, s), 3.36-3.55 (3H, m), 3.75 (3H, s), 3.92 (1H, m), 3.97 (2H, s), 6.93 (1H, t), 7.07 (1H, m), 7.17 (1H, m), 8.50 (1H, s); M/z (LC-MS, ESI+), RT=1.89 minute (M+H=431.19).

(b) 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5,7-dimethyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (58)

(0.021ml 0.28mmol) joins 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-2, and (0.108g is in acetate 0.25mmol) (4ml) solution for 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (57) with hydrazine hydrate.The solution that obtains was at room temperature stirred 2 days.The mixture that obtains is evaporated to drying, and, obtains raw product resistates and methylbenzene azeotropic, with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance (0.033g, productive rate 31.9%). ¹H NMR (399.902MHz, DMSO) δ 1.30 (1H, m), 1.44 (1H, m), 1.72 (1H, m), 1.87 (1H, m), 2.27 (3H, s), 2.51 (3H, s), 3.05 (1H, m), 3.26 (3H, s), 3.32 (2H, m), 3.42 (1H, m), 3.92 (1H, m), 4.08 (2H, s), 7.14-7.30 (3H, m), 11.08 (1H, s), 11.94 (1H, s); M/z (LC-MS, ESI+), RT=1.44 minute (M+H=413.36).

(c) 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-1,2,5-trimethylammonium-1H-pyrroles-3-carboxylate methyl ester (59)

With methyl iodide (0.029ml; 0.46mmol) join 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-2; 5-dimethyl-1H-pyrroles-3-carboxylate methyl ester (57) (0.100g; 0.23mmol) and salt of wormwood (0.048g is in DMF 0.35mmol) (5ml) solution.The suspension that obtains was at room temperature stirred 16 hours.Unreacted is complete, and (0.029ml 0.46mmol) and with suspension at room temperature further stirred 4 hours to add methyl iodide again.Unreacted is complete, therefore temperature is increased to 50 ℃, and reaction mixture was further stirred 3 hours.Reaction mixture is poured in the water (75ml), and (3 * 50ml) extractions, water (50ml) and saturated brine (50ml) clean the organic layer that merges with EtOAc.With organic layer is passed through Na ₂SO ₄Drying is filtered and evaporation, obtains raw product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be colourless jelly (0.082g, productive rate 79%); M/z (LC-MS, ESI+), RT=2.06 minute (M+H=445.30).

(d) 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5,6,7-trimethylammonium-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (60)

(0.014ml 0.19mmol) joins 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-1,2, and (0.076g is in acetate 0.17mmol) (4ml) solution for 5-trimethylammonium-1H-pyrroles-3-carboxylate methyl ester (59) with hydrazine hydrate.The solution that obtains was at room temperature stirred 2 days.The mixture that obtains is evaporated to drying, and, obtains raw product resistates and methylbenzene azeotropic, with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (4.00mg, productive rate 5.49%); M/z (LC-MS, ESI+), RT=1.60 minute (M+H=427.11).

Embodiment 28

(a) 5-methyl isophthalic acid H-pyrroles-3-carboxylate methyl ester (61)

Under nitrogen atmosphere; with methyl acrylate (0.523ml; 5.81mmol) and 1-(1-isocyano-ethylsulfonyl)-4-methylbenzene (1.215g; 5.81mmol) diethyl ether (16.5ml) and DMSO (8.5ml) solution dropwise join the sodium hydride (60% dispersion) that is stirring (0.372g be in diethyl ether 9.29mmol) (25ml) suspension.The suspension that obtains was at room temperature stirred 2 hours.Reaction mixture is poured in 2% sodium chloride solution (200ml), separated each layer, and use Et ₂(4 * 50ml) extraction water solution merge organic layer to O, and water (50ml) cleans, and passes through Na ₂SO ₄Drying is filtered and evaporation, obtains light brown solid.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-40% ethyl acetate.Pure fraction is evaporated to drying, obtains required compound, be white solid (0.527g, productive rate 65.2%); ¹H NMR (400.132MHz, CDCl ₃) δ 2.19 (3H, s), 3.72 (3H, s), 6.23 (1H, m), 7.21 (1H, m), 8.09 (1H, s); M/z (LC-MS, ESI+), RT=1.30 minute (not detecting a large amount of ions).

(b) 4-(2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-5-methyl isophthalic acid H-pyrroles-3-carboxylate methyl ester (62)

Under 0 ℃, nitrogen atmosphere, (0.104g, anhydrous DCM (5ml) solution 0.74mmol) dropwise join the aluminum chloride that is stirring, and (0.226g is in anhydrous DCM (10ml) suspension 1.69mmol) with 5-methyl isophthalic acid H-pyrroles-3-carboxylate methyl ester (61).The suspension that obtains was stirred 10 minutes down at 0 ℃.In the described suspension that is stirring, dropwise add under 0 ℃ essential acyl chloride solution [before the preparation; Under 0 ℃ with 1-chloro-N, N, (0.090ml 0.68mmol) joins 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) acetate (43) (0.200g to 2-trimethylammonium third-1-alkene-1-amine, 0.68mmol) anhydrous DCM (5ml) solution in, and stirred 2 hours].The suspension that obtains is warming up to room temperature, and at room temperature further stirs 18 hours.React incomplete, (0.226g 1.69mmol), and at room temperature further stirred solution 1 hour to add aluminum chloride again.Reaction mixture is poured among ice (50ml) and the dense HCl (2ml).Separate each layer, and with DCM (2 * 50ml) aqueous layer extracted.Use successively 2M HCl (2 * 75ml), water (50ml) and saturated NaHCO ₃(50ml) clean the organism that merges.Organic layer is passed through Na ₂SO ₄Drying is filtered and evaporation, obtains rough product.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100% ethyl acetate.Pure fraction is evaporated to drying, obtains desired substance, be colourless jelly (0.117g, productive rate 41.5%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.44-1.77 (3H, m), 1.85 (1H, m), 2.10 (3H, s), 3.08 (1H, m), 3.29 (3H, s), 3.36-3.55 (3H, m), 3.75 (3H, s), 3.92 (1H, m), 4.16 (2H, m), 6.93 (1H, t), 7.07-7.12 (2H, m), 7.21 (1H, m), 8.82 (1H, s); M/z (LC-MS, ESI+), RT=1.72 minute (M+H=417.20).

(c) 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5-methyl-2,6-dihydro-1H-pyrroles [3,4-d] pyridazine-1-ketone (63)

(0.023ml, (2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) ethanoyl)-(0.117g is in acetate 0.28mmol) (4ml) for 5-methyl isophthalic acid H-pyrroles-3-carboxylate methyl ester (62) 0.31mmol) to join 4-with hydrazine hydrate.The solution that obtains was at room temperature stirred 2 days.The mixture that obtains is evaporated to drying, and, obtains raw product resistates and methylbenzene azeotropic, with preparation HPLC (Waters XBridgePrep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (0.022g, productive rate 19.65%); ¹H NMR (400.132MHz, DMSO) δ 1.21 (1H, m), 1.34 (1H, m), 1.62 (1H, m), 1.77 (1H, m), 2.25 (3H, s), 2.96 (1H, m), 3.15-3.36 (6H, m), 3.82 (1H, s), 4.05 (2H, s), 7.07-7.15 (2H, m), 7.19 (1H, m), 7.33 (1H, s), 11.22 (1H, s), 12.15 (1H, s); M/z (LC-MS, ESI+), RT=1.39 minute (M+H=399.15).

Embodiment 29

(a) (Z)-5,6-two chloro-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji) isobenzofuran-1 (3H)-ketone (64)

To be equipped with 5,6-dichloro isobenzofuran-1, and the 3-diketone (370mg, 1.71mmol), 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) acetate (43) (496mg, 1.68mmol) and sodium acetate (34mg, flask 0.41mmol) are placed in 210 ℃ of pre-heated aluminium blocks.The mixture that obtains is further heated under 240 ℃ and stirred 1 hour, then cooling.With resistates ethanol (～use ultrasonication in 25ml), and collect solid by suction filtration, obtain raw product (128mg, 16.92%), it directly uses and need not to be further purified; M/z (LC-MS, ESI+), RT=2.79 (M+H 450.1 ﹠amp; 452.0).

(b) 6,7-two chloro-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (65)

With (Z)-5, (127mg 0.28mmol) puts into water (2ml) and N, in the dinethylformamide (0.500ml) to isobenzofuran-1 (3H)-ketone (64) to 6-two chloro-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji).(0.080ml 1.65mmol) handles the mixture that obtains, and reaction is heated to 105 ℃ of stirrings 90 minutes with hydrazine hydrate.With reaction mixture cooling, (～10ml) dilution and collect solid by suction filtration obtains raw product to water, with quick silicon-dioxide chromatogram purification raw product, uses the pure ethyl acetate wash-out.Pure fraction is evaporated to drying, and resistates is pulverized in diethyl ether, dry under vacuum, obtain desired substance, be white solid (46.0mg, productive rate 35%); ¹H NMR (400.132MHz, DMSO) δ 1.30-1.40 (1H, m), 1.44-1.53 (1H, m), 1.72-1.79 (1H, m), and 1.87-1.94 (1H, m), 3.04-3.11 (1H, m), 3.29 (3H, s), 3.30-3.40 (2H, m), 3.44-3.49 (1H, m), 3.92-4.00 (1H, m), 4.37 (2H, s), 7.26 (1H, t), 7.38-7.45 (2H, m), 8.29 (1H, s), 8.40 (1H, s), 12.83 (1H, s); M/z (LC-MS, ESI+), RT=2.13 (M+H 464 ﹠amp; 465.9).

Embodiment 30

(a) (Z)-6-chloro-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji) isobenzofuran-1 (3H)-ketone (67) and (Z)-5-chloro-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji) isobenzofuran-1 (3H)-ketone (66)

5-chlorine isobenzofuran-1 will be housed, 3-diketone (313mg, 1.71mmol), 2-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) phenyl) acetate (43) (496mg, 1.68mmol) and sodium acetate (37mg, flask 0.45mmol) are placed in 210 ℃ of pre-heated aluminium blocks.The mixture that obtains is further heated under 240 ℃ and stirred 45 minutes, then cooling.With resistates put into ethanol (～10ml) and add diethyl ether (～40ml).Collect the solid of gained by suction filtration.With filtrate evaporation, and with quick silicon-dioxide residue purified by chromatography, gradient is the isohexane solution of 50-100%EtOAc.The required compound fraction is evaporated to drying, obtains 2 kinds of regional isomers 67, be light brown solid (27.0mg, 3.87%); ¹HNMR (400.132MHz, DMSO) δ 1.38-1.55 (2H, m), 1.79-1.86 (1H, m), 1.89-1.96 (1H, m), 3.12-3.19 (1H, m), 3.28 (3H, s), 3.36-3.43 (2H, m), 3.45-3.51 (1H, m), 3.96-4.03 (1H, m), 7.05 (1H, s), 7.45 (1H, t), 7.80 (1H, dd), 7.88-7.92 (1H, m), 7.98 (1H, dd), 8.07 (1H, d), 8.12 (1H, d); M/z (LC-MS, ESI+), RT=2.65 (M+H416.1 ﹠amp; 417.9); With 66 be light brown solid (55.0mg, 7.87%); ¹H NMR (400.132MHz, DMSO) δ 1.38-1.54 (2H, m), 1.79-1.85 (1H, m), 1.89-1.96 (1H, m), 3.11-3.19 (1H, m), 3.28 (3H, s), 3.35-3.43 (2H, m), 3.44-3.50 (1H, m), 3.95-4.02 (1H, m), 7.09 (1H, s), 7.44 (1H, t), 7.73 (1H, dd), 7.77 (1H, dd), 7.85-7.89 (1H, m), 7.99 (1H, d), 8.28 (1H, d); M/z (LC-MS, ESI+), RT=2.63 (M+H 416.1 ﹠amp; 417.9) it directly used in next stage.

(b) 6-chloro-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (68)

With (Z)-5-chloro-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji) isobenzofuran-1 (3H)-ketone (66) (53mg, 0.13mmol) be suspended in water (1.5ml) and N, in the dinethylformamide (0.5ml), and with a hydrazine hydrate (35 μ l 0.72mmol) handle.The mixture that obtains was stirred 1 hour down at 105 ℃.(～5ml) dilution is with DCM (2 * 5ml) extraction mixtures with reaction mixture cooling and water.The organism that merges is evaporated to drying, obtains raw product, with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of acetonitrile as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (13mg, productive rate 24%); ¹H NMR (400.132MHz, DMSO) δ 1.28-1.38 (1H, m), 1.40-1.50 (1H, m), 1.49-1.76 (1H, m), and 1.83-1.91 (1H, m), 3.01-3.09 (1H, m), 3.25 (3H, s), and 3.26-3.37 (2H, m), 3.40-3.46 (1H, m), 3.88-3.96 (1H, m), 4.34 (2H, s), 7.22 (1H, t), 7.35-7.42 (2H, m), 7.87 (1H, dd), 8.02 (1H, d), 8.26 (1H, d), 12.68 (1H, s); M/z (LC-MS, ESI+), RT=2.24 (M+H 430.6).

(c) 7-chloro-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (69)

With (Z)-6-chloro-3-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) Ben Yajiaji) isobenzofuran-1 (3H)-ketone (67) (25mg, 0.06mmol) place water (1ml) and N, in the dinethylformamide (0.5ml), and with a hydrazine hydrate (20 μ l 0.41mmol) handle.The mixture that obtains was stirred 1 hour down at 105 ℃.With the reaction mixture cooling, and water (～5ml) dilute, with DCM (2 * 5ml) extraction mixtures.The organism that merges is evaporated to drying, obtains raw product, with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of acetonitrile as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white foam (10mg, productive rate 39%); ¹H NMR (400.132MHz, DMSO) δ 1.26-1.35 (1H, m), 1.40-1.49 (1H, m), 1.67-1.74 (1H, m), and 1.83-1.90 (1H, m), 2.99-3.06 (1H, m), 3.26 (3H, s), and 3.26-3.36 (2H, m), 3.39-3.45 (1H, m), 3.88-3.95 (1H, m), 4.33 (2H, s), 7.22 (1H, t), 7.31 (1H, dd), 7.38-7.42 (1H, m), 7.94 (1H, dd), 8.00 (1H, d), 8.21 (1H, d), 12.72 (1H, s); M/z (LC-MS, ESI+), RT=2.23 (M+H 430.7).

Embodiment 31

(a) 4-(3-(4-tert.-butoxy piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (70)

With triethylamine (0.750ml, 5.38mmol) and O-benzotriazole-1-base-N, N, N ', (1.15g 3.03mmol) handles 2-fluoro-5-((4-oxygen-3 to N '-tetramethyl-urea hexafluorophosphate, 4-dihydro phthalazines-1-yl) phenylformic acid (1) (650mg methyl), 2.18mmol) and 4-tert.-butoxy piperidines (350mg, N,N-dimethylacetamide 2.23mmol) (11ml) solution.The mixture that obtains at room temperature stirred spend the night.With reaction mixture pour into water (～200ml) in, collect the solid of gained by suction filtration, and dry, obtain raw product, with quick silicon-dioxide chromatogram purification raw product, gradient is the dichloromethane solution of 2-10% methyl alcohol.Pure fraction is evaporated to drying, and dry under vacuum, obtain desired substance, be white solid (460mg, productive rate 48.2%); ¹H NMR (400.132MHz, DMSO) δ 1.14 (9H, s), 1.19-1.38 (2H, m), 1.53-1.60 (1H, m), and 1.70-1.77 (1H, m), 3.02-3.10 (1H, m), 3.16-3.28 (2H, m), and 3.70-3.77 (1H, m), 4.02-4.10 (1H, m), 4.33 (2H, s), 7.20 (1H, t), 7.31-7.35 (1H, m), 7.38-7.42 (1H, m), 7.81-7.90 (2H, m), 7.97 (1H, d), 8.27 (1H, dd), 12.56 (1H, s); M/z (LC-MS, ESI+), RT=2.13 (M+H 438.2).

Embodiment 32

(a) oxygen base 4-[[3-[4-[(2-methyl-prop-2-yl)] piperidines-1-carbonyl] phenyl] methyl]-2H-phthalazines-1-ketone (71)

In the 20ml bottle, add 3-[(4-oxygen-3H-phthalazines-1-yl) methyl] phenylformic acid (1b) (50mg, 0.178mmol).In this bottle, add N,N-dimethylacetamide (2ml) and triethylamine (70 μ l, 0.445mmol).This mixture was stirred 5 minutes, add O-benzotriazole-1-base-N then, N, N ', N '-tetramethyl-urea hexafluorophosphate further stirs this solution 5 minutes.Add 4-[(2-methyl-prop-2-yl then) the oxygen base] piperidine hydrochlorate (35mg, (1ml) solution of N,N-dimethylacetamide 0.178mmol) and triethylamine (25 μ l, 0.179mmol), to react and at room temperature stir 2 hours, use the preparation HPLC purifying then, obtain required compound; M/z (LC-MS, ESI+), RT=1.74 (M+H 420.4).

Embodiment 33

(a) 4-[[4-fluoro-3-[4-(3-picoline-2-yl) oxygen phenylpiperidines-1-carbonyl] phenyl] methyl]-2H-phthalazines-1-ketone (72a)

(60% mineral oil disperses thing with sodium hydride; 3.8mg, 0.157mmol) processing rough 4-[[4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) phenyl] methyl]-2H-phthalazines-1-ketone (2a) (50mg, N,N-dimethylacetamide 0.131mmol) (1ml) solution, and will react at room temperature and stir, stop up to effervesce.(16mg 0.144mmol), and is sealed in reaction mixture in the microwave tube, is heated to 170 ℃, keeps 250 seconds under this temperature, is cooled to room temperature then to add 2-fluoro-3-picoline then.Use this mixture of preparation HPLC purifying then, obtain desired substance; M/z (LC-MS, ESI+), RT=4.22 (M+H 473.2)

(b) similar embodiment

Use the method similar methods of describing with 72a, make 4-[[4-fluoro-3-(4-hydroxy piperidine-1-carbonyl) phenyl] methyl]-2H-phthalazines-1-ketone reacted 10 minutes at 170 ℃ with suitable fluorine pyridine, used the preparation HPLC purifying, obtained required compound.

Embodiment 34

(a) 4-((2-(4-tert.-butoxy piperidines-1-carbonyl) pyridin-4-yl) methyl) phthalazines-1 (2H)-ketone (73)

Under 25 ℃, with 4-tert.-butoxy piperidine hydrochlorate (227mg, 1.17mmol) and triethylamine (0.327ml, 2.35mmol) N, disposable 4-((the 4-oxygen-3 that is stirring that joins of dinethylformamide (3ml) solution, 4-dihydro phthalazines-1-yl) pyridine carboxylic acid (5) (300mg methyl), 1.07mmol), triethylamine (0.327ml, 2.35mmol) and O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (607mg, 1.60mmol) N, in dinethylformamide (3ml) solution.The solution that obtains was stirred 4 hours down at 25 ℃, use preparation HPLC (Waters XBridge Prep C18OBD post, 5 μ silicon-dioxide, diameter 30mm then, long 100mm) purification of crude mixture, water (containing 1%NH3) that reduces gradually with polarity and the mixture of MeCN are as elutriant.The fraction that will contain required compound is evaporated to dry and freeze-drying, obtains required compound, is solid (250mg, productive rate, 55.7%); ¹H NMR (400.132MHz, DMSO) δ 1.15 (9H, s), 1.25-1.40 (2H, m), 1.56-1.63 (1H, m), and 1.71-1.79 (1H, m), 3.08-3.23 (2H, m), 3.44-3.50 (1H, m), and 3.71-3.78 (1H, m), 4.01-4.08 (1H, m), 4.40 (2H, s), 7.39 (1H, dd), 7.48-7.49 (1H, m), 7.83-7.87 (1H, m), 7.90 (1H, td), 7.94-7.97 (1H, m), 8.28 (1H, dd), 8.46-8.48 (1H, m), 12.60 (1H, s); M/z (LC-MS, ESI+), RT=1.67 (M+H 421.5).

Embodiment 35

(a) 4-(4-fluoro-3-(4-hydroxy-4-methyl piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (74)

Under 25 ℃, air atmosphere, with O-benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea hexafluorophosphate (572mg, 1.51mmol) disposable 2-fluoro-5-((the 4-oxygen-3 that joins, 4-dihydro phthalazines-1-yl) phenylformic acid (1) (300mg methyl), 1.01mmol) and triethylamine (0.308ml, N 2.21mmol) is in dinethylformamide (3ml) solution.The solution that obtains was stirred 10 minutes at 25 ℃.Dropwise add 4-methyl piperidine-4-alcohol hydrochloride (154mg, 1.02mmol) and triethylamine (0.308ml, N 2.21mmol), dinethylformamide (1ml) solution, and the solution that obtains stirred 2 hours at 25 ℃.With DCM (100ml) diluted reaction mixture, and successively water (3 * 50ml) and saturated brine (20ml) clean.Pass through MgSO ₄Dry organic layer filters and evaporation, obtains raw product, and with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is jelly (94mg, productive rate 23.63%); ¹H NMR (400.132MHz, DMSO) δ 1.13 (3H, s), 1.29-1.47 (4H, m), 3.20-3.28 (4H, m), 4.32 (2H, s), 4.41 (1H, s), 7.19 (1H, d), 7.31 (1H, dd), 7.38-7.42 (1H, m), 7.83 (1H, td), 7.88 (1H, td), 7.96 (1H, d), 8.27 (1H, dd), 12.56 (1H, s); M/z (LC-MS, ESI+), RT=1.45 (M+H 396.4).

Embodiment 36

(a) 4-(2-morpholino-2-oxygen oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (75)

In toluene (75ml), add 4-hydroxy piperidine-1-carboxylic acid tert-butyl ester (10g, 49.69mmol), 4-butyl ammonium hydrogen sulfate (0.844g, 2.48mmol) and 2-chloro-1-morpholino ethyl ketone (8.13g, 49.69mmol), (40g, water 400.03mmol) (45ml) solution will be reflected at 25 ℃ of following stirrings and spend the night to wherein adding NaOH.Reaction mixture water (100ml) finishes reaction, uses Et ₂(MgSO is passed through in 3 * 75ml) extractions to O ₄Dry organic layer filters and evaporation, obtains white solid.This white solid is dissolved in DCM and evaporation carefully again, obtains yellow jelly, add diethyl ether, add isohexane then, up to observing muddy solution.System is stirred, obtain solid, collect this solid by filtering, and dry under vacuum, obtain required compound, be white solid (15.30g, productive rate 94%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.45 (9H, s), 1.58-1.49 (0H, m), 1.72-1.61 (2H, m), 1.89-1.82 (2H, m), 3.12-3.06 (2H, m), 3.61-3.54 (6H, m), 3.68-3.67 (4H, m), 3.80-3.72 (1H, m), 4.18 (2H, s).

(b) 1-morpholino-2-(piperidin-4-yl oxygen base) ethyl ketone (76)

(5g, (30ml 180.00mmol) in the solution, will be reflected at 25 ℃ and stir 2 hours down 15.23mmol) to join the propan-2-ol of 6.0HCl with 4-(2-morpholino-2-oxygen oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (75).Reaction mixture is evaporated to drying, uses the ion-exchange chromatogram purification crude material, use the SCX post.Use 7M NH ₃/ MeOH is evaporated to drying with required product wash-out from post with fraction, obtains yellow jelly.By distillation purifying raw product under 0.72mBar, be collected in 155 ℃ of distillatory cuts, obtain desired substance, be colourless jelly (1.950g, productive rate 56.1%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.50-1.41 (2H, m), 1.96-1.89 (2H, m), 2.64-2.58 (2H, m), 3.10-3.04 (2H, m), 3.50-3.43 (1H, m), 3.63-3.56 (4H, m), 3.70-3.67 (4H, m), 4.17 (2H, s); The NH proton disappears.

(c) 4-(4-fluoro-3-(4-(2-morpholino-2-oxygen oxyethyl group) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (77)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (0.2g, 0.67mmol), 1-morpholino-2-(piperidin-4-yl oxygen base) ethyl ketone (76) (0.153g, 0.67mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.215g 0.67mmol) is dissolved in DMF (10ml) to 3-tetramethyl-isourea a tetrafluoro borate, adds DIPEA (0.117ml in this solution, 0.67mmol), and will react and stir 1 hour.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white foam (0.214g, 62.8%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.64-1.53 (1H, m), 1.75-1.67 (2H, m), 1.86-1.75 (1H, m), 1.98-1.93 (1H, m), 3.16-3.07 (1H, m), 3.57-3.39 (3H, m), 3.64-3.58 (2H, m), 3.72-3.66 (5H, m), 4.11-3.99 (1H, m), 4.19 (2H, q), 4.27 (2H, s), 7.02 (1H, t), 7.32-7.26 (2H, m), 7.77-7.69 (3H, m), 8.49-8.44 (1H, m), 10.51 (1H, s); Owing to have rotational isomer, therefore have very wide peak; M/z (LC-MS, ESI+), RT=1.48 (M+H 509).

(d) 4-(2-morpholino oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (78)

With 4-(2-morpholino-2 oxygen oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (75) (8.84g, 26.92mmol) be dissolved in exsiccant THF (50ml), in this solution, add borine-methyl thioether complex compound (20.19ml, 40.38mmol), will be reflected at 40 ℃ of following stirrings and at room temperature spend the night then in 3 hours.Colloidal mixture is evaporated, and finish reaction with 2.0N yellow soda ash (50ml), (MgSO is passed through in 3 * 75ml) extractions with EtOAc ₄Dry organic layer filters and evaporation, obtains desired substance, is colourless liquid (6.50g, productive rate 77%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.45 (9H, s), 1.54-1.45 (2H, m), 1.84-1.79 (2H, m), 2.90-2.84 (2H, m), 3.16-3.03 (6H, m), 3.79-3.67 (5H, m), 3.97 (2H, t), 4.15-4.10 (2H, m).

(e) 4-(2-(piperidin-4-yl oxygen base) ethyl) morpholine (79)

(7.0g, (30ml 180.00mmol) in the solution, will be reflected at 25 ℃ and stir 2 hours down 22.26mmol) to join the propan-2-ol of 6.0HCl with 4-(2-morpholino oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (78).Reaction mixture is evaporated to drying, and be dissolved in methyl alcohol again (75ml) and water (7ml), by stirring this solution with solid sodium hydroxide alkalization 1 hour.To react and filter and evaporation, obtain colloidal solid.This solid was stirred 20 minutes in ethyl acetate (75ml), filter and evaporation, obtain limpid liquid.By distillation purifying raw product under 1.2mBar, be collected in 120 ℃ of distillatory cuts, obtain desired substance, be colourless oily matter (3.10g, 65.0%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.47-1.38 (2H, m), 1.93-1.89 (2H, m), 2.53-2.51 (4H, m), 2.63-2.56 (4H, m), 3.08 (2H, dt), 3.38-3.31 (1H, m), 3.61 (2H, t), 3.71 (4H, dd).

(f) 4-(4-fluoro-3-(4-(2-morpholino oxyethyl group) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (80)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (0.2g, 0.67mmol), 4-(2-(piperidin-4-yl oxygen base) ethyl) morpholine (79) (0.144g, 0.67mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.215g 0.67mmol) is dissolved in DMF (10ml) to 3-tetramethyl-isourea a tetrafluoro borate, adds DIPEA (0.117ml in this solution, 0.67mmol), and will react and stir 1 hour.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white foam (0.055g, productive rate 16.6%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.60-1.48 (1H, m), 1.83-1.64 (2H, m), 1.94-1.88 (1H, m), 2.59-2.57 (4H, m), 2.64 (2H, t), 3.14-3.05 (1H, m), 3.49-3.37 (1H, m), 3.58-3.51 (1H, m), 3.66-3.60 (3H, m), 3.74-3.72 (4H, m), 4.06-3.94 (1H, m), 4.28 (2H, s), 7.01 (1H, t), 7.32-7.26 (2H, m), 7.77-7.70 (3H, m), 8.48-8.44 (1H, m), 10.77 (1H, s); M/z (LC-MS, ESI+), RT=1.57 (M+H 495).

Embodiment 37

(a) 4-(2-(dimethylamino) 2-oxygen oxyethyl group) piperidines-1-carboxylic acid benzene methyl (81)

In toluene (50ml), add 4-hydroxy piperidine-1-carboxylic acid benzene methyl (5g, 21.25mmol), 4-butyl ammonium hydrogen sulfate (0.361g, 1.06mmol) and 2-chloro-N, N-N,N-DIMETHYLACETAMIDE (4.13g, 27.63mmol), and (21g, water 21.02mmol) (30ml) solution will be reflected at 25 ℃ of following stirrings and spend the night to add sodium hydroxide in this solution.Water (100ml) finishes reaction with reaction mixture, uses Et ₂(MgSO is passed through in 3 * 75ml) extractions to O ₄Dry organic layer filters and evaporation, obtains required compound, is orange liquid (6.00g, productive rate 88%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.65-1.55 (2H, m), 1.93-1.86 (2H, m), 2.95 (3H, s), 3.03 (3H, s), 3.25-3.18 (2H, m), 3.63-3.57 (1H, m), 3.85-3.79 (2H, m), 4.17 (2H, s), 5.12 (2H, s), 7.38-7.30 (5H, m).

(b) N, N-dimethyl-2-(piperidin-4-yl oxygen base) ethanamide (82)

In ethanol (40ml), add 4-(2-(dimethylamino)-2-oxygen oxyethyl group) piperidines-1-carboxylic acid benzene methyl (81) (3.0g, 9.36mmol) and palladium-carbon catalyst (0.100g, 0.94mmol).It was stirred 3 hours under nitrogen atmosphere.To react filtration, and, obtain the clarifying oily matter of viscosity solvent evaporation.By distillation purifying raw product under 0.5mBar, be collected in 90 ℃ of distillatory cuts, obtain desired substance, be colorless oil (1.000g, productive rate 57.3%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.51-1.42 (2H, m), 1.98-1.92 (2H, m), 2.63-2.57 (2H, m), 2.95 (3H, s), 3.10-3.05 (5H, m), 3.51-3.44 (1H, m), 4.17 (2H, s) NH proton disappearances.

(c) 2-(1-(2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzoyl) piperidin-4-yl oxygen base)-N,N-dimethylacetamide (83)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (0.2g, 0.67mmol), N, N-dimethyl-2-(piperidin-4-yl oxygen base) ethanamide (82) (0.125g, 0.67mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea a tetrafluoro borate (0.215g, 0.67mmol) be dissolved in DMF (10ml), (0.117ml 0.67mmol), and will react and stir 1 hour to add DIPEA in this solution.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is Off-white solid (0.191g, productive rate 61.1%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.98-1.55 (4H, m), 2.96 (3H, s), 3.03 (3H, s), and 3.17-3.06 (1H, m), 3.60-3.43 (2H, m), 3.72-3.67 (1H, m), 4.08-3.99 (1H, m), 4.19 (2H, q), 4.28 (2H, s), 7.01 (1H, t), 7.32-7.26 (2H, m), 7.78-7.70 (3H, m), 8.48-8.46 (1H, m), 10.89 (1H, s); M/z (LC-MS, ESI+), RT=1.55 (M+H 467).

(d) 4-(2-(dimethylamino) oxyethyl group) piperidines-1-carboxylic acid benzene methyl (84)

With 4-(2-(dimethylamino)-2-oxygen oxyethyl group) piperidines-1-carboxylic acid benzene methyl (81) (4.5g, 14.50mmol) be dissolved in exsiccant THF (50ml), in this solution, add borine-methyl thioether complex compound (10.53ml, 21.07mmol), will be reflected at 40 ℃ of following stirrings and at room temperature spend the night then in 3 hours.With the colloidal mixture evaporation, and use 2.0N Na ₂CO ₃(50ml) finish reaction, (MgSO is passed through in 3 * 75ml) extractions with EtOAc ₄Dry organic layer filters and evaporation, obtains desired substance, is colourless liquid (4.00g, productive rate 93%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.60-1.51 (2H, m), 1.88-1.78 (2H, m), 2.65 (6H, s), 2.97 (2H, t), 3.29-3.22 (2H, m), 3.54-3.48 (1H, m), 3.79-3.73 (2H, m), 3.85 (2H, t), 5.12 (2H, s), 7.36-7.28 (5H, m).

(e) N, N-dimethyl-2-(piperidin-4-yl oxygen base) ethamine (85)

Under 50 ℃, the nitrogen atmosphere of 5bar, with 4-(2-(dimethylamino) oxyethyl group) piperidines-1-carboxylic acid benzene methyl (84) (4.5g, 14.69mmol) and 5% palladium-carbon catalyst (JM 87L type; 0.9g, EtOH 0.21mmol) (50ml) solution stirring 3 hours.To react filtration and, obtain yellow jelly solvent evaporation.This jelly is dissolved in diethyl ether (50ml) and filtration.Remove and desolvate, obtain orange,, be collected in 70 ℃ of distillatory cuts, obtain desired substance, be colourless liquid (1.100g, 55.9%) by this oily matter of distillation purifying under 1.0mBar; ¹H NMR (400.132MHz, CDCl ₃) δ 1.47-1.38 (2H, m), 1.61 (1H, s), 1.93-1.89 (2H, m), 2.27 (6H, s), 2.50 (2H, t), 2.63-2.56 (2H, m), 3.11-3.05 (2H, m), 3.38-3.31 (1H, m), 3.56 (2H, t).

(f) 4-(3-(4-(2-(dimethylamino) oxyethyl group) piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (86)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (0.2g, 0.67mmol), N, N-dimethyl-2-(piperidin-4-yl oxygen base) ethamine (85) (0.116g, 0.67mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea a tetrafluoro borate (0.215g, 0.67mmol) be dissolved in DMF (10ml), (0.117ml 0.67mmol), and will react and stir 1 hour to add DIPEA in this solution.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (0.087g, productive rate 28.7%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.62-1.51 (1H, m), 1.84-1.66 (3H, m), 1.95-1.89 (1H, m), 2.28 (6H, s), 2.52 (2H, t), 3.15-3.01 (1H, m), 3.48-3.40 (1H, m), 3.62-3.51 (3H, m), 4.07-3.98 (1H, m), 4.27 (2H, s), 7.01 (1H, t), 7.32-7.25 (2H, m), 7.77-7.70 (3H, m), 8.47-8.45 (1H, m), 10.62 (1H, s); M/z (LC-MS, ESI+), RT=1.67 (M+H 452).

Embodiment 38

(a) 4-(2-(3,3-difluoro azetidine-1-yl)-2-oxygen oxyethyl group) piperidines-1-carboxylic acid benzene methyl (87)

With 2-(1-(benzyl oxygen base carbonyl) piperidin-4-yl oxygen base) acetate (4.12g, 14.05mmol), 3,3-difluoro azetidine hydrochloride (1.4g, 10.81mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (3.47g 10.81mmol) is dissolved in DMF (50ml) to 3-tetramethyl-isourea a tetrafluoro borate, adds DIPEA (4.91ml in this solution, 28.10mmol), and will react to stir and spend the night.With solvent evaporation, and make reaction mixture finish reaction, use Et with 2M NaOH (50ml) ₂(MgSO is passed through in 3 * 50ml) extractions to O ₄Dry organic layer filters and evaporation, obtains desired substance, is yellow jelly (2.59g, productive rate 65.1%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.67-1.49 (2H, m), 1.90-1.81 (2H, m), 3.26-3.19 (2H, m), 3.58-3.51 (1H, m), 3.84-3.78 (2H, m), 4.13 (2H, s), 4.36 (2H, t), 4.60 (2H, t), 5.12 (2H, s), 7.40-7.29 (5H, m).

(b) 1-(3,3-difluoro azetidine-1-yl)-2-(piperidin-4-yl oxygen base) ethyl ketone (88)

In ethanol (40ml), add 4-(2-(3,3-difluoro azetidine-1-yl)-2-oxygen oxyethyl group) piperidines-1-carboxylic acid benzene methyl (87) (2.59g, 7.03mmol) and palladium-carbon catalyst (0.075g, 0.70mmol).With this solution at H ₂(1.417g 703.09mmol) stirred 3 hours under the atmosphere.To react and filter and evaporating solvent, obtain viscosity clarification oily matter.By distillation purifying raw product under 0.5mBar, be collected in 90 ℃ of distillatory cuts, obtain desired substance, be colorless oil (0.540g, productive rate 32.8%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.49-1.40 (2H, m), 1.94-1.89 (2H, m), 2.65-2.59 (2H, m), 3.10-3.05 (2H, m), 3.44-3.38 (1H, m), 4.13 (2H, s), 4.35 (2H, t), 4.64 (2H, t); NH disappears

(c) (((2-(3 for 4-for 3-for 4-; 3-difluoro azetidine-1-yl)-and 2-oxygen oxyethyl group) piperidines-1-carbonyl)-the 4-luorobenzyl) phthalazines-1 (2H)-ketone (89) and 2-(1-(2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzoyl) piperidin-4-yl oxygen base) acetate (90)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1) (0.2g, 0.67mmol), 1-(3,3-difluoro azetidine-1-yl)-2-(piperidin-4-yl oxygen base) ethyl ketone (88) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.215g 0.67mmol) is dissolved in DMF (10ml) to 3-tetramethyl-isourea a tetrafluoro borate, adds DIPEA (0.117ml in this solution, 0.67mmol), and will react and stir 1 hour.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly.With preparation HPLC (WatersXBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains 4-(3-(4-(2-(3,3-difluoro azetidine-1-yl)-2-oxygen oxyethyl group) piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (89), is white solid (0.061g, productive rate 17.7%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.67-1.48 (2H, m), 1.86-1.75 (1H, m), 2.01-1.89 (1H, m), 3.19-3.06 (1H, m), 3.56-3.39 (2H, m), 3.66-3.57 (1H, m), 4.17-3.95 (3H, m), 4.28 (2H, s), 4.39-4.33 (2H, m), 4.62-4.57 (2H, m), 7.02 (1H, t), 7.31-7.26 (2H, m), 7.79-7.70 (3H, m), 8.48-8.46 (1H, m), 10.53 (1H, s) m/z (LC-MS, ESI+), RT=1.83 (M+H 515).And 2-(1-(2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzoyl) piperidin-4-yl oxygen base) acetate (90), be white solid (0.016g, 5.50%); 1H NMR (400.132MHz, CDCl3) δ 1.68-1.60 (1H, m), 1.84-1.71 (2H, m), 2.05-1.96 (1H, m), 3.16-3.04 (1H, m), 3.53-3.45 (2H, m), and 3.78-3.68 (1H, m), 4.23-4.04 (3H, m), 4.32-4.28 (2H, m), 7.07-7.02 (1H, m), 7.30-7.24 (2H, m), 7.86-7.74 (3H, m), 8.47-8.42 (1H, m), 11.12 (1H, s) COOH disappears; M/z (LC-MS, ESI+), RT=0.91 (M+H 440).

Embodiment 39

(a) (S)-4-(1-methoxy propyl-2-base oxygen base) pyridine (91)

In THF (250ml), add pyridine-4-alcohol (10g, 105.15mmol), (R)-1-methoxyl group propan-2-ol (9.48g, 105.15mmol) and triphenylphosphine (30.3g 115.67mmol), and stirred 10 minutes.(22.49ml 115.67mmol), and will stir 1 hour under will being reflected at 25 ℃ slowly to add DIAD in this solution.Evaporating solvent also adds diethyl ether (100ml).In this solution, add a small amount of triphenylphosphine oxidation thing, and will react and stir 20 minutes, obtain solid, solid is abandoned.Evaporating solvent, and, use Et with light yellow gluey thing 2.0HCl acidifying ₂O (alkalize the aqueous solution with solid KOH then by 1 * 75ml) extraction.Use Et then ₂(3 * 75ml) extract this aqueous solution to O, pass through MgSO ₄Dry organic layer filters and evaporation, obtains yellow jelly.Be purified by under 0.43mBar, distilling, be collected in 80 ℃ of distillatory cuts, obtain desired substance, be colorless oil (15.30g, productive rate 87%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.34 (3H, d), 3.40 (3H, s), 3.50 (1H, dd), 3.58 (1H, dd), 4.68-4.60 (1H, m), 6.82 (2H, d), 8.40 (2H, d); M/z (LC-MS, ESI+), RT=1.28 (M+H 168).

(b) (S)-4-(1-methoxy propyl-2-base oxygen base) piperidines (92)

With (S)-4-(1-methoxy propyl-2-base oxygen base) pyridine (91) (15g, 89.71mmol) and the stirring 16 hours under 80 ℃, the nitrogen atmosphere of 50bar of MeOH (50ml) solution of platinum oxide (IV).Therefore analysis revealed does not react, and adds 20ml acetate, and temperature is risen to 100 ℃, pressure rises to 80bar, and restir 18 hours is still only observed very a spot of reaction.The 5% rhodium aluminum oxide that adds five equilibrium, and with mixture heated overnight under 100 ℃, 80bar.GCMS-2 shows that more product forms, and therefore reaction is replaced, and proceeds under 100 ℃, as if impurity peaks no longer increases simultaneously.GCMS-3 shows that more product forms, and therefore reaction is replaced, and proceeds 48 hours under 100 ℃, as if impurity peaks no longer increases simultaneously.GCMS-4 shows that more product forms, and therefore reaction is replaced, and proceeds 48 hours under 100 ℃, as if impurity peaks no longer increases simultaneously.GCMS-5 shows the SM that trace is only arranged, and main peak is a product, and impurity has increase slightly.To react filtration, and evaporating solvent, with 2M NaOH (75ml) reaction is finished, (MgSO is passed through in 3 * 75ml) extractions with EtOAc ₄Dry organic layer filters and evaporation, obtains yellow liquid.By distillation purifying raw product under 0.89mBar, be collected in 70 ℃ of distillatory cuts, obtain required compound, be colourless liquid (2.500g, productive rate 16.08%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.14 (3H, d), 1.47-1.37 (2H, m), 1.94-1.83 (2H, m), 2.62-2.56 (2H, m), 3.10-3.06 (2H, m), 3.30-3.26 (1H, m), 3.40-3.36 (4H, m), 3.50-3.44 (1H, m), 3.75-3.66 (1H, m); NH disappears.

(S)-4-(4-fluoro-3-(4-(1-methoxy propyl-2-base oxygen base) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (93)

With 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (1) (0.2g, 0.67mmol), (S)-4-(1-methoxy propyl-2-base oxygen base) piperidines (92) (0.151g, 0.87mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.280g 0.87mmol) is dissolved among the DMF (10ml) 3-tetramethyl-isourea a tetrafluoro borate, adds DIPEA (0.152ml in this solution, 0.87mmol), and will react and stir 1 hour.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly, and with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white foam; (0.215g, productive rate 70.7%) ¹H NMR (400.132MHz, CDCl ₃) δ 1.13 (3H, t), 1.61-1.49 (1H, m), 1.78-1.65 (2H, m), 1.91-1.87 (1H, m), 3.17-3.01 (1H, m), 3.31-3.27 (1H, m), 3.39-3.35 (4H, m), 3.57-3.45 (2H, m), 3.72-3.67 (2H, m), 4.06-3.97 (1H, m), 4.27 (2H, s), 7.01 (1H, t), 7.31-7.26 (2H, m), 7.79-7.70 (3H, m), 8.49-8.45 (1H, m), 10.50 (1H, s); M/z (LC-MS, ESI+), RT=1.81 (M+H 454).

Embodiment 40

(a) 3,5-dibromo isobenzofuran-1 (3H)-ketone (94)

With 5-bromine isobenzofuran-1 (3H)-ketone (E)-2,2 '-(diazene-1,2-two bases) two (2-methyl propionitrile) (0.540g, 3.29mmol) with 1-bromine tetramethyleneimine-2, (5.85g 32.86mmol) is dissolved in CCl to the 5-diketone ₄, and reflux 2 hours.To react cooling and filtration, evaporated filtrate obtains desired substance, is yellow solid (6.05g, productive rate 63.1%) that it directly used in next stage.

(b) (6-bromo-3-oxygen-1,3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (95)

With 3,5-dibromo isobenzofuran-1 (3H)-ketone (94) (6.2g, 21.24mmol) and triphenylphosphine (5.57g, 21.24mmol) reflux is spent the night in tetrahydrofuran (THF) (100ml).To react cooling and filtration, obtain desired substance, be white solid (7.20g, productive rate 71.5%); ¹H NMR (400.132MHz, CDCl ₃) δ 7.23 (1H, s), 7.59 (1H, d), 7.72-7.67 (7H, m), 7.92-7.84 (9H, m), 10.20 (1H, s);

(c) 5-((7-bromo-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorine benzonitrile (96)

With (6-bromo-3-oxygen-1; 3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (95) (9.0g; 16.24mmol) and 2-fluoro-5-formyl radical benzonitrile (2.91g; 19.49mmol) be dissolved in DCM (60ml); (2.94ml 21.11mmol) also will react stirring and spend the night to add triethylamine in this solution.Reaction mixture water (50ml) finishes reaction, and (MgSO is passed through in 2 * 75ml) extractions with DCM ₄Dry organic layer filters and evaporation, obtains orange jelly.Make it by the silicon-dioxide plug, use eluent ethyl acetate, obtain yellow jelly.To wherein adding entry (40ml), EtOH (40ml) and DMF (4ml).(8.13g 162.39mmol) also will react reflux and spend the night to add hydrazine hydrate.To react cooling,, clean with EtOH (25ml) by filtering collecting precipitation, and dry air, obtain required compound, be white solid (2.410g, productive rate 41.4%) that it need not to be further purified in use; ¹H NMR (400.132MHz, DMSO) δ 4.38 (2H, s), 7.48 (1H, t), 7.75-7.71 (1H, m), 7.89 (1H, dd), 8.02 (1H, d), 8.18 (1H, d), 8.22 (1H, s), 12.64 (1H, s); M/z (LC-MS, ESI-), RT=2.18 (M-H 358).

(d) 5-((7-bromo-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorobenzoic acid (97)

In ethanol (20ml) and water (80ml), add 5-((7-bromo-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorine benzonitrile (96) (2.4g, 6.70mmol) and potassium hydroxide (3.76g 67.01mmol), and heated 5 hours down at 100 ℃.Evaporate ethanol, and with ethyl acetate (1 * 75ml) extraction water solution.Use then dense HCl with acidified aqueous solution to pH1, obtain solid, with solid filtering, water cleans and is dry, obtains desired substance, is light brown solid.(2.000g, productive rate 79%); ¹H NMR (400.132MHz, DMSO) δ 4.37 (2H, s), 7.26 (1H, t), 7.61-7.57 (1H, m), 7.83-7.82 (1H, m), 8.01 (1H, d), 8.18 (1H, d), 8.22 (1H, s), 12.67 (1H, s), 13.19 (1H, s); M/z (LC-MS, ESI+), RT=0.88 (M+H 376).

(e) 6-bromo-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (98)

With 5-((7-bromo-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorobenzoic acid (97) (1.0g, 2.65mmol), 4-methoxyl group piperidines (0.336g, 2.92mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3, (0.936g 2.92mmol) is dissolved in DMF (40ml) to 3-tetramethyl-isourea a tetrafluoro borate, adds DIPEA (0.509ml in this solution, 2.92mmol), and will react and stir 1 hour.Evaporation DMF, and 2M NaOH (75ml) the end reaction of rough jelly, (3 * 75ml) extractions, organic layer passes through MgSO with EtOAc ₄Drying is filtered and evaporation, obtains brown jelly.Make this jelly by the silicon-dioxide plug, use eluent ethyl acetate, obtain white solid, the major part of these white solids need not to be further purified in use.With a sample (100mg) preparation HPLC (Waters XBridgePrep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (66mg); ¹H NMR (400.132MHz, CDCl ₃) δ 1.62-1.48 (1H, m), 1.85-1.68 (2H, m), 1.94-1.89 (1H, m), 3.18-3.07 (1H, m), 3.35 (3H, s), 3.47-3.40 (2H, m), 3.67-3.53 (1H, m), 4.06-3.89 (1H, m), 4.23 (2H, s), 7.04 (1H, t), 7.31-7.26 (2H, m), 7.86-7.84 (2H, m), 8.33-8.31 (1H, m), 10.55 (1H, s); M/z (LC-MS, ESI+), RT=1.94 (M+H 476).

(f) 6-amino-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (99)

In diox (15ml), add 6-bromo-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (98) (0.2g, 0.42mmol), diphenylmethyl ketoimine (0.076g, 0.42mmol) and cesium carbonate (0.275g, 0.84mmol), and make the degassing of this system with nitrogen.(0.019g, 0.08mmol) (0.073g 0.13mmol), will be reflected at 95 ℃ and heat 2 hours down with (9,9-dimethyl-9H-xanthene-4,5-two bases) two (diphenylphosphines) to add palladium diacetate in this system.Reaction mixture water (50ml) finishes reaction, and (MgSO is passed through in 3 * 50ml) extractions with EtOAc ₄Dry organic layer filters and evaporation, obtains brown jelly.It was stirred 1 hour in 2.0N HCl (5ml), and reaction mixture finishes reaction with 2M NaOH (10ml), and (MgSO is passed through in 3 * 15ml) extractions with EtOAc ₄Dry organic layer filters and evaporation, obtains brown jelly.With preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purification of crude product, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (0.024g, 13.87%); ¹H NMR (500.133MHz, DMSO) δ 1.47-1.38 (2H, m), 1.82-1.73 (2H, m), 2.89 (3H, s), 3.23-3.16 (2H, m), 3.27 (2H, s), 3.46-3.41 (1H, m), 4.13 (2H, s), 5.78 (2H, s), 6.87 (1H, d), 7.00 (1H, dd), 7.14 (1H, t), 7.22 (1H, dd), 7.38-7.34 (1H, m), 7.93 (1H, d), 11.60 (1H, s); M/z (LC-MS, ESI+), RT=1.56 (M+H 411).

Embodiment 41

(a) 3-bromo-4-chlorine isobenzofuran-1 (3H)-ketone (100)

(10g, 59.32mmol), (E)-2,2 '-(diazene-1,2-two bases) two (2-methyl propionitrile) (0.974g, 5.93mmol) and 1-bromine tetramethyleneimine-2, (11.61g 65.25mmol) is dissolved in CCl to the 5-diketone with 4-chlorine isobenzofuran-1 (3H)-ketone ₄(100ml), and reflux 2 hours.To react cooling and filtration,, obtain required compound, be yellow jelly (14.20g, productive rate 97%) the filtrate evaporation; ¹H NMR (400.132MHz, CDCl ₃) δ 7.32 (1H, s), 7.61 (1H, t), 7.73 (1H, d), 7.88 (1H, d); M/z (LC-MS, ESI+), RT=2.28 (not detecting M+H).

(b) (7-chloro-3-oxygen-1,3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (101)

With 3-bromo-4-chlorine isobenzofuran-1 (3H)-ketone (100) (14g, 56.57mmol) and triphenylphosphine (14.84g 56.57mmol) is dissolved in THF (200ml), and reflux 2 hours.To react cooling and filtration, evaporated filtrate obtains required compound, is yellow jelly (22.5g, productive rate 78%); ¹H NMR (400.132MHz, CDCl ₃) δ 7.54-7.51 (2H, m), 7.65-7.59 (7H, m), 7.79-7.74 (3H, m), 8.07-8.02 (6H, m), 10.41 (1H, s); M/z (LC-MS, ESI+), RT=2.09 (not detecting M+H).

(c) 5-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) 2-fluorine benzonitrile (102)

With (7-chloro-3-oxygen-1; 3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (101) (9.3g; 21.64mmol) and 2-fluoro-5-formyl radical benzonitrile (3.55g; 23.80mmol) be dissolved in DCM (60ml); in this solution, add triethylamine (3.92ml; 28.13mmol), and will react to stir and spend the night.Evaporation reaction mixture obtains brown solid.In this solid, add entry (40ml), EtOH (40ml) and DMF (4ml).(10.83g 216.35mmol), and will react reflux and spend the night to add hydrazine hydrate.To react cooling and filter the collecting precipitation thing, clean with EtOH (25ml), and dry air, obtain required compound, be yellow solid (6.10g, 90%) that it need not to be further purified in use; ¹H NMR (400.132MHz, DMSO) δ 4.64 (2H, s), 7.44 (1H, t), 7.63-7.59 (1H, m), 7.78 (1H, dd), 7.81 (1H, t), 7.98 (1H, dd), 8.33 (1H, dd); M/z (LC-MS, ESI+), RT=2.17 (M-H 312).

(d) 5-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorobenzoic acid (103)

In ethanol (30ml) and water (70ml), add 5-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorine benzonitrile (102) (6.1g, 19.44mmol) and potassium hydroxide (10.91g 194.44mmol), and heated 5 hours down at 100 ℃.Evaporate ethanol, with ethyl acetate (1 * 75ml) extraction water solution.Use then dense HCl with acidified aqueous solution to pH1, obtain solid, with solid filtering, water cleans and is dry, obtains required compound, is light brown solid (5.73g, 89%); ¹H NMR (400.132MHz, DMSO) δ 4.63 (2H, s), 7.21 (1H, dd), 7.44-7.40 (1H, m), 7.64 (1H, dd), 7.82 (1H, t), 7.98 (1H, dd), 8.34 (1H, dd), 12.87 (1H, s), 13.12 (1H, s); M/z (LC-MS, ESI+), RT=0.87 (M+H 333).

(e) 5-chloro-4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (104a)

With 5-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorobenzoic acid (103) (0.2g, 0.60mmol), 4-methoxyl group piperidines (0.069g, 0.60mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea a tetrafluoro borate (0.251g, 0.78mmol) be dissolved in DMF (10ml), (0.136ml 0.78mmol), and will react and stir 1 hour to add DIPEA in this solution.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly, and with preparation HPLC (Waters XBridge PrepC18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white foam (0.100g, 38.7%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.60-1.49 (1H, m), 1.83-1.64 (2H, m), 1.94-1.89 (1H, m), 3.17-3.05 (1H, m), 3.35 (3H, s), 3.48-3.44 (2H, m), 3.61-3.48 (1H, m), 4.03-3.93 (1H, m), 4.65 (2H, s), 7.00 (1H, t), 7.17-7.14 (2H, m), 7.66 (1H, t), 7.80 (1H, dd), 8.49 (1H, dd) NH disappears; M/z (LC-MS, ESI+), RT=1.89 (M+H 430).

(f) 5-chloro-4-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (104b)

With 5-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-2-fluorobenzoic acid (103) (0.2g, 0.60mmol), 4-oxyethyl group piperidines (0.078g, 0.60mmol) and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea a tetrafluoro borate (0.251g, 0.78mmol) be dissolved in DMF (10ml), (0.136ml 0.78mmol), and will react and stir 1 hour to add DIPEA in this solution.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly, and with preparation HPLC (Waters XBridge PrepC18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white foam (0.052g, 19.49%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.20 (3H, t), 1.59-1.48 (1H, m), 1.81-1.63 (2H, m), and 1.94-1.89 (1H, m), 3.16-3.07 (1H, m), 3.57-3.45 (4H, m), 4.09-4.00 (1H, m), 4.65 (2H, s), 6.99 (1H, t), 7.17-7.13 (2H, m), 7.26 (1H, s), 7.66 (1H, t), 7.80 (1H, dd), 8.49 (1H, dd); NH disappears; M/z (LC-MS, ESI+), RT=2.08 (M+H 444).

Embodiment 42

(a) 4-(2-oxygen-2-(tetramethyleneimine-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (105)

In toluene (50ml), add 4-hydroxy piperidine-1-carboxylic acid tert-butyl ester (4g, 19.87mmol), 4-butyl ammonium hydrogen sulfate (0.337g, 0.99mmol) and 2-chloro-1-tetramethyleneimine-1-base-ethyl ketone (3.87g, 25.84mmol), in this solution, add NaOH (19.87g, water 198.75mmol) (20ml) solution, and will be reflected at 25 ℃ and stir down and spend the night.Reaction mixture water (100ml) finishes reaction, uses Et ₂(3 * 75ml) extractions, organic layer passes through MgSO to O ₄Drying is filtered and evaporation, obtains desired substance, is orange liquid (7.20g, productive rate＞100%); ¹H NMR (400.13MHz, DMSO-d ₆) δ 1.40 (9H, s), 1.86 (8H, m), 3.02 (4H, s), 3.38 (2H, t), 3.54 (1H, m), 3.60 (2H, m), 4.07 (2H, s).

(b) 2-(piperidin-4-yl oxygen base)-1-(tetramethyleneimine-1-yl) ethyl ketone (106)

With 4N HCl De diox (3ml, 12mmol) solution joins 4-(2-oxygen-2-(tetramethyleneimine-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (105) (3.5g, 11.20mmol) methyl alcohol (3ml) solution in, reaction mixture was at room temperature stirred 6 hours.Remove and desolvate, obtain desired substance, be its hydrochloride form.This salt is dissolved in methyl alcohol (50ml) and add MP-carbonate (14.88g, 33.61mmol).The mixture that obtains is filtered and removes and desolvate, obtain desired substance (1.700g, productive rate 71.5%); ¹H NMR (400.13MHz, DMSO-d ₆) δ 1.34 (2H, m), 1.73 (2H, q), 1.86 (4H, q), 2.43 (2H, m), 2.94 (2H, m), 3.39 (4H, t), 4.04 (2H, s).

(c) 4-(4-fluoro-3-(4-(2-oxygen-2-(tetramethyleneimine-1-yl) oxyethyl group) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (107)

In DMA (4ml), add 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1) (0.20g, 0.67mmol) and HBTU (0.381g, 1.01mmol), in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.179ml, 1.01mmol), add then 2-(piperidin-4-yl oxygen base)-1-(tetramethyleneimine-1-yl) ethyl ketone (106) (0.142g, 0.67mmol).To react and stir 2 hours, be evaporated to drying then, and with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 21mm, long 100mm) purifying, water (containing 1%NH+) that reduces gradually with polarity and the mixture of MeCN are as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white solid (0.168g, 50.9%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.90-1.56 (m, 6H), 2.01-1.93 (m, 2H), 3.16-3.07 (m, 1H), and 3.57-3.42 (m, 6H), 3.72 (septet, J=3.6Hz, 1H), and 4.06-4.00 (m, 1H), 4.12 (q, J=11.9Hz, 2H), 4.26 (s, 2H), 7.02 (t, J=8.8Hz, 1H), and 7.32-7.25 (m, 2H), 7.79-7.70 (m, 3H), 8.47-8.44 (m, 1H), 10.01-9.97 (m, 1H); M/z (LC-MS, ESI+), RT=1.66 (M+H493).

(d) 4-(2-(tetramethyleneimine-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (108)

With 4-(2-oxygen-2-(tetramethyleneimine-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (105) (3.5g, 11.20mmol) be dissolved in exsiccant THF (50ml), in this solution, add borine-methyl thioether complex compound (8.40ml, 16.81mmol), will be reflected at 40 ℃ of following stirrings and at room temperature spend the night then in 3 hours.Colloidal mixture is evaporated, and finish reaction with 2.0N yellow soda ash (50ml), (3 * 75ml) extractions, organic layer passes through MgSO with EtOAc ₄Drying is filtered and evaporation, obtains desired substance, is colourless liquid, and it directly used in next stage; ¹H NMR (400.132MHz, CDCl ₃) δ 1.38-1.58 (m, 9H), 1.73-1.86 (m, 4H), 1.86-1.97 (m, 2H), 2.09-2.21 (m, 2H), 2.79-2.91 (m, 2H), 2.96 (t, 2H), 3.06-3.17 (m, 2H), 3.18-3.27 (m, 2H), 3.49 (septet, 2H), 3.65-3.76 (m, 2H), 3.92 (t, 1H).

(e) 1-(2-(piperidin-4-yl oxygen base) ethyl) piperidines (109)

4N HCl De diox (6ml) solution is joined 4-(2-(tetramethyleneimine-1-yl) oxyethyl group), and (6.5g in MeOH 20.80mmol) (16ml) solution, at room temperature stirred reaction mixture 6 hours piperidines-1-carboxylic acid tert-butyl ester (108).Remove and desolvate, obtain required compound, be its hydrochloride form.This salt is dissolved in methyl alcohol (50ml) and add MP-carbonate (14.88g, 33.61mmol).The mixture that obtains is filtered and removes and desolvate, obtain desired substance, be clarifying liquid, it need not to be further purified in use; ¹H NMR (400.132MHz, CDCl ₃) δ 1.33-1.47 (2H, m), 1.50-1.71 (3H, m), 1.86-1.96 (4H, m), 2.50-2.67 (6H, m), 3.00-3.14 (4H, m), 3.34 (1H, septets), 3.55 (2H, t), 3.60 (2H, t).

(f) 4-(4-fluoro-3-(4-(2-(tetramethyleneimine-1-yl) oxyethyl group) piperidines-1-carbonyl) benzyl) phthalazines 1 (2H)-ketone (110)

In DMF (4ml), add 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1) (0.20g, 0.67mmol) and HBTU (0.381g, 1.01mmol), in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.179ml, 1.01mmol), add then 4-(2-(tetramethyleneimine-1-base oxygen base) ethyl) piperidines (109) (0.133g, 0.67mmol).To react and stir 2 hours, and be evaporated to drying then, and with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white solid (0.287g, productive rate 89%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.65-1.81 (6H, m), 1.87-1.96 (3H, m), 2.53-2.61 (3H, m), and 2.65-2.72 (2H, m), 3.04-3.14 (1H, m), 3.38-3.57 (3H, m), 3.61 (2H, t), 3.97-4.06 (1H, m), 4.28 (2H, s), 7.01 (1H, t), 7.21-7.33 (2H, m), 7.68-7.80 (3H, m), 8.43-8.48 (1H, m), 9.85-9.95 (1H, m); M/z (LC-MS, ESI+), RT=1.88 (M+H 479.5).

Embodiment 43

(a) 4-(2-oxygen-2-(piperidines-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (111)

In toluene (75ml), add 4-hydroxy piperidine-1-carboxylic acid tert-butyl ester (10g, 49.69mmol), 4-butyl ammonium hydrogen sulfate (0.844g, 2.48mmol) and 2-chloro-1-(piperidines-1-yl) ethyl ketone (8.08g, 49.69mmol), in this solution, add sodium hydroxide (49.7g, 496.86mmol) water (20ml) solution, and will be reflected at 25 ℃ and stir down and spend the night.Reaction mixture water (100ml) finishes reaction, uses Et ₂(MgSO is passed through in 3 * 75ml) extractions to O ₄Dry organic layer filters and evaporation, obtains raw product, is orange liquid (15.60g, productive rate 96%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.49 (9H, s), 1.51-1.60 (2H, m), 1.61-1.68 (2H, m), 1.81-1.90 (3H, m), and 3.05-3.13 (2H, m), 3.35-3.47 (2H, m), 3.50-3.62 (4H, m), 3.71-3.79 (2H, m), 3.80-3.89 (2H, m), 4.13-4.19 (2H, m).

(b) 1-(piperidines-1-yl)-2-(piperidin-4-yl oxygen base) ethyl ketone (112)

4N HCl De diox (6ml) solution is joined 4-(2-oxygen-2-(piperidines-1-yl) oxyethyl group), and (7.5g in MeOH 22.98mmol) (20ml) solution, at room temperature stirred reaction mixture 6 hours piperidines-1-carboxylic acid tert-butyl ester (111).Remove and desolvate, obtain the hydrochloride (2.300g, 44.2%) of 1-(piperidines-1-yl)-2-(piperidin-4-yl oxygen base) ethyl ketone.Then this salt is dissolved in methyl alcohol (50ml) and add MP-carbonate (14.88g, 33.61mmol).The mixture that obtains is filtered and removes and desolvate, obtain required compound (2.300g, 44.2%), it need not to be further purified in use.

(c) 4-(4-fluoro-3-(4-(2-oxygen-2-(piperidines-1-yl) oxyethyl group) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (113)

In DMA (4ml), add 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1) (0.20g, 0.67mmol) and HBTU (0.381g, 1.01mmol), in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.179ml, 1.01mmol), add then 1-(piperidines-1-yl)-2-(piperidin-4-yl oxygen base) ethyl ketone (112) (0.152g, 0.67mmol).To react and stir 2 hours, and be evaporated to drying then, and with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance (0.303g, productive rate 89%); ¹H NMR (400.132MHz, DMSO) δ 1.07-1.39 (8H, m), 1.46-1.56 (1H, m), 1.61-1.71 (1H, m), and 2.76-2.85 (1H, m), 3.02-3.09 (2H, m), 3.10-3.20 (4H, m), and 3.34-3.41 (1H, m), 3.66-3.75 (1H, m), 3.90-3.93 (2H, m), 4.08-4.12 (2H, m), 6.99 (1H, t), 7.09-7.21 (2H, m), 7.57-7.69 (1H, m), 7.72-7.76 (1H, m), 8.02-8.06 (2H, m), 12.34 (1H, s); M/z (LC-MS, ESI+), RT=1.76 (M+H 507.5).

(d) 4-(2-(piperidines-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (114)

With 4-(2-oxygen-2-(piperidines-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (111) (7.5g, 22.98mmol) be dissolved in exsiccant THF (100ml), in this solution, add borine-methyl thioether complex compound (17.23ml, 34.46mmol), will be reflected at 40 ℃ of following stirrings and at room temperature spend the night then in 3 hours.Colloidal mixture is evaporated, and finish reaction with 2.0N yellow soda ash (50ml), (3 * 75ml) extractions, organic layer passes through MgSO with EtOAc ₄Drying is filtered and evaporation, obtains 4-(2-(piperidines-1-yl) oxyethyl group) piperidines-1-carboxylic acid tert-butyl ester (7.3g, productive rate 95%), is colourless liquid, and it need not to be further purified in use.

(e) 1-(2-(piperidin-4-yl oxygen base) ethyl) piperidines (115)

4N HCl De diox (6ml) solution is joined 4-(2-(piperidines-1-yl) oxyethyl group), and (6.5g in MeOH 20.80mmol) (16ml) solution, at room temperature stirred reaction mixture 6 hours piperidines-1-carboxylic acid tert-butyl ester (114).Remove and desolvate, obtain the hydrochloride (0.540gg, 12.22%) of 1-(2-(piperidin-4-yl oxygen base) ethyl) piperidines.Then this salt is dissolved in methyl alcohol (50ml) and add MP-carbonate (14.88g, 33.61mmol).The mixture that obtains is filtered and removes and desolvate, obtain desired substance, be colourless liquid, it need not to be further purified in use; ¹H NMR (400.132MHz, CDCl ₃) δ 1.33-1.47 (2H, m), 1.50-1.71 (3H, m), 1.86-1.96 (4H, m), 2.50-2.67 (6H, m), 3.00-3.14 (4H, m), 3.34 (1H, septets), 3.55 (2H, t), 3.60 (2H, t).

(f) 4-(4-fluoro-3-(4-(2-(piperidines-1-yl) oxyethyl group) piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (116)

In DMA (4ml), add 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1) (0.20g, 0.67mmol) and HBTU (0.381g, 1.01mmol), in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.179ml, 1.01mmol), add then 1-(2-(piperidin-4-yl oxygen base) ethyl) piperidines (115) (0.214g, 1.01mmol).To react and stir 2 hours, be evaporated to drying then, and with preparation HPLC (Waters XBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, water (containing 1%NH3) that reduces gradually with polarity and the mixture of MeCN are as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound (0.069g, productive rate 20.74%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.36-1.45 (2H, m), 1.64-1.94 (8H, m), 2.37-2.50 (4H, m), 2.55 (2H, t), 3.03-3.15 (1H, m), 3.37-3.49 (1H, m), 3.50-3.65 (4H, m), 3.92-4.04 (1H, m), 4.30 (2H, s), 7.00 (1H, t), 7.21-7.32 (2H, m), 7.68-7.79 (3H, m), 8.43-8.49 (1H, m), 10.49 (1H, s); M/z (LC-MS, ESI+), RT=2.01 (M+H 493.5).

Embodiment 44

(a) 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzonitrile (117)

With (7-chloro-3-oxygen-1; 3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (101) (10g, 23.26mmol) (3.20g 24.43mmol) is dissolved in DCM (60ml) with 3-formyl radical benzonitrile; (4.22ml 30.24mmol) also will react stirring and spend the night to add triethylamine in this solution.Reaction mixture is filtered, obtain white solid.In this solid, add entry (40ml), EtOH (40ml) and DMF (4ml).(11.65g 232.64mmol) also will react reflux and spend the night to add hydrazine hydrate.To react cooling,, clean with EtOH (25ml) by filtering the collecting precipitation thing, and dry air, obtain desired substance, be white solid (3.75g, productive rate 54.5%) that it need not to be further purified in use; ¹H NMR (400.132MHz, DMSO) δ 4.66 (2H, s), 7.53-7.47 (2H, m), 7.68-7.66 (2H, m), 7.81 (1H, t), 7.97 (1H, d), 8.33 (1H, d), 12.55 (1H, s); M/z (LC-MS, ESI-), RT=2.04 (M-H 294).

(b) 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (118)

In ethanol (30ml) and water (70ml), add 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzonitrile (117) (3.75g, 12.68mmol) and potassium hydroxide (7.11g 126.81mmol), and heated 5 hours down at 100 ℃.Evaporate ethanol, with ethyl acetate (1 * 75ml) extraction water solution.Use then dense HCl with acidified aqueous solution to pH1, obtain solid, with solid filtering, water cleans and is dry, obtains desired substance, is light brown solid (3.81g, productive rate 95%); ¹H NMR (400.132MHz, DMSO) δ 4.67 (2H, s), 7.42-7.41 (2H, m), 7.72 (1H, s), 7.79-7.76 (1H, m), 7.81 (1H, t), 7.97 (1H, dd), 8.34 (1H, dd), 12.88 (1H, s); M/z (LC-MS, ESI+), RT=0.92 (M+H 315).

(c) 5-chloro-4-(3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (119a)

In DMF (5ml), add 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) phenylformic acid (118) (200mg methyl), 0.64mmol) and 4-methoxyl group piperidines (73.2mg, 0.64mmol), in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.170ml, 0.95mmol), add then HBTU (362mg, 0.95mmol).Reaction mixture was stirred 4 hours down at 0 ℃.With preparation type LCMS (WatersXBridge Prep C18 OBD post, 5 μ silicon-dioxide, diameter 30mm, long 100mm) purification of crude mixture, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white crystalline solid (62.2mg, productive rate 23.76%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.62-2.00 (3H, m), 3.02-3.29 (2H, m), 3.35 (3H, s), 3.37-3.65 (3H, m), 3.87-4.08 (1H, m), 4.69 (2H, s), 7.15-7.24 (3H, m), 7.29-7.34 (1H, m), 7.65 (1H, t), 7.77-7.81 (1H, m), 8.48 (1H, m), 9.85-10.01 (1H, m); M/z (LC-MS, ESI+), RT=1.80 (M+H 412.1).

(d) 5-chloro-4-(3-(4-oxyethyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (119bd)

In DMF (5ml), add 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) phenylformic acid (118) (200mg methyl), 0.64mmol) and 4-oxyethyl group piperidines, in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.170ml, 0.95mmol), add then HBTU (362mg, 0.95mmol).Remove and to desolvate, and raw product is dissolved in acetonitrile, and with preparation HPLC (Waters XBridge Prep C18OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains desired substance, is white solid (111mg, productive rate 41.0%); ¹H NMR (400.132MHz, DMSO) δ 1.11 (3H, t), 1.17-1.46 (2H, m), 1.58-1.92 (2H, m), 2.97-3.42 (4H, m), 3.42-3.55 (3H, m), 4.68 (2H, s), 7.13 (1H, s), 7.21 (2H, t), 7.35 (1H, t), 7.81 (1H, t), 7.94-7.99 (1H, m), 8.32-8.36 (1H, m), 12.91 (1H, s); M/z (LC-MS, ESI+), RT=1.96 (M+H 426.7)

Embodiment 45

(a) 4-methoxyl group isobenzofuran-1 (3H)-ketone (121)

Under 21 ℃, air atmosphere, with methyl iodide (1.313ml, 20.78mmol) join 4-hydroxyl isobenzofuran-1 (3H)-ketone (3.12g, 20.78mmol) and salt of wormwood (5.74g is in DMF 41.56mmol) (50ml) solution.The solution stirring that obtains is spent the night.Remove and desolvate, obtain yellow solid, then it is finished reaction with salt of wormwood (70ml).With ethyl acetate (3 * 75ml) extraction mixtures.The extraction liquid that evaporation merges obtains desired substance, is yellow solid (3.18g, productive rate 93%) that it need not to be further purified in use; ¹H NMR (400.132MHz, CDCl ₃) δ 3.30 (2H, s), 5.37 (2H, s), 7.31-7.42 (2H, m), 7.51-7.61 (1H, m).

(b) 3-bromo-4-methoxyl group isobenzofuran-1 (3H)-ketone (122)

(3.18g, 19.37mmol) with 1-bromine tetramethyleneimine-2, (3.62g 20.34mmol) is dissolved in tetracol phenixin (40ml) to the 5-diketone, and reflux with 4-methoxyl group isobenzofuran-1 (3H)-ketone (121).Add (E)-2, (0.318g 1.94mmol), and will stir under will being reflected at 80 ℃ and spends the night 2 '-(diazene-1,2-two bases) two (2-methyl propionitrile).To react cooling and filtration.Remove and desolvate, obtain desired substance, be orange solids (3.15g, 66.9%) that it need not to be further purified in use; ¹H NMR (400.132MHz, DMSO) δ 3.91 (3H, s), 6.66 (1H, s), 7.35-7.40 (2H, m), 7.57-7.65 (1H, m).

(c) (7-methoxyl group-3-oxygen-1,3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (123)

With 3-bromo-4-methoxyl group isobenzofuran-1 (3H)-ketone (122) (3.15g, 12.96mmol) and triphenylphosphine (3.4g 12.96mmol) is dissolved in THF (50ml), and reflux whole weekend.To react cooling and filtration, obtain desired substance, be white solid (3.50g, 53.4%); ¹H NMR (400.132MHz, CDCl ₃) δ 3.42 (3H, s), 7.10 (1H, d), 7.22-7.28 (1H, m), 7.47-7.53 (1H, m), 7.59-7.65 (6H, m), 7.74-7.80 (3H, m), 7.91-8.00 (6H, m), 9.80 (1H, s).

(d) 2-fluoro-5-((8-methoxyl group-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzonitrile (124)

With (7-methoxyl group-3-oxygen-1; 3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (123) (3.5g; 6.93mmol) and 2-fluoro-5-formyl radical benzonitrile (1.033g; 6.93mmol) be dissolved in DCM (30ml); in this solution, add triethylamine (1.255ml; 9.00mmol), and will react to stir and spend the night.Remove and desolvate, obtain white solid.In this solid, add entry (20ml), EtOH (20ml) and DMF (2ml).(3.36ml 69.26mmol), and will react reflux and spend the night to add hydrazine hydrate.To react cooling and, use EtOH (25ml) to clean by filtering the collecting precipitation thing, and dry air, obtain desired substance, be white solid (2.70g,＞100%) that it need not to be further purified in use; ¹H NMR (400.132MHz, DMSO) δ 3.79 (3H, s), 4.40 (2H, s), 7.31-7.39 (1H, m), 7.47-7.59 (2H, m), 7.66-7.73 (2H, m), 7.78-7.82 (1H, m), 12.40-12.62 (1H, m).

(e) 2-fluoro-5-((8-methoxyl group-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (125)

In ethanol (20ml) and water (50ml), add 2-fluoro-5-((8-methoxyl group-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) benzonitrile (124) (2.7g, 8.73mmol) and potassium hydroxide (4.90g 87.30mmol), and heated 5 hours at 100 ℃.Evaporate ethanol, and with ethyl acetate (1 * 75ml) extraction water solution.Use then dense HCl with acidified aqueous solution to pH1, obtain solid, with its filtration, water cleans and is dry, obtains desired substance, is white solid (2.370g, 83%); ¹H NMR (400.132MHz, DMSO) δ 3.82 (3H, s), 4.40 (2H, s), 7.16-7.23 (1H, m), 7.40-7.46 (2H, m), 7.64-7.68 (1H, m), 7.76 (1H, t), 7.84-7.88 (1H, m), 12.96-13.28 (1H, m), 12.59-12.63 (1H, m).

(f) 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl)-5-methoxyl group phthalazines-1 (2H)-ketone (126a)

In DMF (5ml), add 2-fluoro-5-((8-methoxyl group-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl) phenylformic acid (125) and 4-methoxyl group piperidines, and adding N-ethyl-N-sec.-propyl third-2-amine in this solution (0.163ml, 0.91mmol), add then HBTU (347mg, 0.91mmol).Remove and desolvate, resistates is dissolved in acetonitrile.Be settled out white solid, it is filtered and drying, obtain desired substance, be white crystalline solid (140mg, productive rate 54.0%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.49-1.60 (1H, m), 1.62-1.83 (2H, m), 1.86-1.96 (1H, m), and 3.07-3.19 (1H, m), 3.32-3.36 (3H, m), 3.41-3.62 (3H, m), 3.78-3.82 (3H, m), 3.94-4.04 (1H, m), and 4.36-4.46 (2H, m), 6.93-7.00 (1H, m), 7.10-7.30 (3H, m), 7.64-7.72 (1H, m), 8.00-8.12 (1H, m), 9.98-10.07 (1H, m); M/z (ES+) (M+H) +=426.4; HPLC RT=1.71 minute.

(g) 4-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-luorobenzyl)-5-methoxyl group phthalazines-1 (2H)-ketone (126b)

In DMF (5ml), add 2-fluoro-5-((8-methoxyl group-4-oxygen-3,4-dihydro phthalazines-1-yl) phenylformic acid (125) (200mg methyl), 0.61mmol) and 4-oxyethyl group piperidines (79mg, 0.61mmol), in this solution, add N-ethyl-N-sec.-propyl third-2-amine (0.163ml, 0.91mmol), add then HBTU (347mg, 0.91mmol).Remove then and desolvate, obtain brown solid, it is dissolved in acetonitrile, be settled out white solid, it is filtered and drying, obtain desired substance, be white crystalline solid (200mg, productive rate 74.7%); ¹H NMR (400.132MHz, CDCl ₃) δ 1.20 (3H, t), 1.46-1.62 (1H, m), 1.63-1.82 (2H, m), and 1.87-1.96 (1H, m), 3.07-3.18 (1H, m), 3.43-3.59 (5H, m), 3.77-3.82 (3H, m), 3.99-4.11 (1H, m), 4.45 (2H, s), 6.91-7.02 (1H, m), 7.11-7.28 (3H, m), 7.62-7.71 (1H, m), 8.03-8.08 (1H, m), 10.29 (1H, s); M/z (ES+) (M+H) +=440.41; HPLC RT=1.87 minute.

Embodiment 46

(a) 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-5-fluorine benzonitrile (127)

With (7-chloro-3-oxygen-1; 3-dihydroisobenzofuran-1-yl) triphenyl bromide phosphine (101) (2.88g; 6.71mmol) and 3-fluoro-5-formyl radical benzonitrile (1.0g; 6.71mmol) be dissolved in DCM (60ml); in this solution, add triethylamine (1.215ml; 8.72mmol), and will react to stir and spend the night.Evaporation reaction mixture obtains brown solid.In this solid, add entry (40ml), EtOH (40ml) and DMF (4ml).(3.36g 67.06mmol), and will react reflux and spend the night to add hydrazine hydrate.To react cooling and filter the collecting precipitation thing, clean with EtOH (25ml), and dry air, obtain required compound, be yellow solid (1.260g, 59.9%) that it need not to be further purified in use; ¹H NMR (400.132MHz, DMSO) δ 4.68 (2H, s), 7.49 (1H, d), 7.58 (1H, s), 7.68 (1H, d), 7.82 (1H, t), 7.99 (1H, dd), 8.34 (1H, dd), 12.82 (1H, s); M/z (LC-MS, ESI-), RT=2.21 (M-H 312).

(b) 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-5-fluorobenzoic acid (128)

In ethanol (30ml) and water (70ml), add 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-5-fluorine benzonitrile (127) (1.26g, 4.02mmol) and potassium hydroxide (2.253g 40.16mmol), and heated 5 hours down at 100 ℃.Evaporate ethanol, and with ethyl acetate (1 * 75ml) extraction water solution.Use then dense HCl with acidified aqueous solution to pH1, obtain solid, with solid filtering, water cleans and is dry, obtains required compound, is light brown solid (1.280g, productive rate 96%); ¹H NMR (400.132MHz, DMSO) δ 4.69 (2H, s), 7.35 (1H, d), 7.50 (1H, d), 7.58 (1H, s), 7.82 (1H, t), 7.99 (1H, d), 8.34 (1H, d), 12.87 (1H, s); M/z (LC-MS, ESI+), RT=0.99 (M+H 333).

(c) 5-chloro-4-(3-(4-oxyethyl group piperidines-1-carbonyl)-5-luorobenzyl) phthalazines-1 (2H)-ketone (129a)

With 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-5-fluorobenzoic acid (128), 4-oxyethyl group piperidines and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea a tetrafluoro borate is dissolved in DMF (10ml), in this solution, add DIPEA, and will react and stir 1 hour.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly, and with preparation HPLC (Waters XBridge PrepC18OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white foam (130mg); ¹H NMR (400.132MHz, CDCl ₃) δ 1.20 (3H, t), 2.00-1.38 (4H, m), 3.29-3.05 (1H, m), 3.57-3.34 (5H, m), 4.08-3.91 (1H, m), 4.68 (2H, s), 6.90 (1H, d), 6.97-6.94 (2H, m), 7.67 (1H, t), 7.80 (1H, d), 8.50 (1H, dd); NH disappears; M/z (LC-MS, ESI+), RT=2.06 (M+H 444).

(d) 5-chloro-4-(3-fluoro-5-(4-methyl piperidine-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (129b)

With 3-((8-chloro-4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-5-fluorobenzoic acid (128), 4-methoxyl group piperidines and 2-(1H-benzo [d] [1,2,3] triazol-1-yl)-1,1,3,3-tetramethyl-isourea a tetrafluoro borate is dissolved in DMF (10ml), in this solution, add DIPEA, and will react and stir 1 hour.Solvent evaporation to dry, is dissolved in acetonitrile (4ml) with jelly, and with preparation HPLC (Waters XBridge PrepC18OBD post, 5 μ silicon-dioxide, diameter 19mm, long 100mm) purifying, the water that reduces gradually with polarity (contains 1%NH ₃) and the mixture of MeCN as elutriant.The fraction that will contain required compound is evaporated to drying, obtains required compound, is white foam (67.0mg); ¹H NMR (400.132MHz, CDCl ₃) δ 1.89-1.40 (4H, m), 3.17-3.06 (1H, m), 3.34 (3H, s), 3.58-3.42 (3H, m), 4.00-3.84 (1H, m), 4.67 (2H, s), 6.97-6.89 (3H, m), 7.67 (1H, t), 7.81 (1H, d), 8.49 (1H, d), 10.29 (1H, s); M/z (LC-MS, ESI+), RT=1.93 (M+H 430).

Embodiment 47

A) resynthesis of 2b

Under 20 ℃, nitrogen atmosphere, with O-benzotriazole-1-base-tetramethyl-urea hexafluorophosphate (45.5g, 119.86mmol) join 2-fluoro-5-((4-oxygen-3 in batches, 4-dihydro phthalazines-1-yl) phenylformic acid (1) (27.5g methyl), 92.20mmol), 4-methoxyl group piperidines (11.68g, 101.42mmol) and triethylamine (30.8ml is in DMA 221.28mmol) (450ml) solution.The solution that obtains was stirred 21 hours down at 20 ℃.Solution is poured in the water (2.5 liters), and, cleaned the extract that merges, dry (MgSO with salt solution (* 3) with EtOAc (* 3) extraction ₄), filter and evaporation, obtain jelly.With quick silicon-dioxide chromatogram purification raw product, gradient is the isohexane solution of 0-100%EtOAc.Pure fraction is evaporated to drying, and uses the EtOAc pulp, obtain 4-(4-fluoro-3-(4-methoxyl group piperidines-1-carbonyl) benzyl) phthalazines-1 (2H)-ketone (2b) (22.45g, 61.6%) after filtration with after the vacuum-drying, be white solid.

B) pulp 2b is with the preparation hydrate

With the concentration of～25mg/ml in water preparation process a) in the suspension of 2b of preparation, and under the steady temperature between 20 to 50 ℃, stirred 48 hours.From water, isolate required hydrate by filtering then.

C) resynthesis of 2f

With triethylamine (31.6ml, 226.89mmol) (27.07g is 90.76mmol) with 4-oxyethyl group piperidines (12.11g to handle 2-fluoro-5-((4-oxygen-3,4-dihydro phthalazines-1-yl) methyl)-phenylformic acid (1), 93.73mmol) N, N-N,N-DIMETHYLACETAMIDE (422ml) solution, and stirred 5 minutes, in 10 minutes time, add O-benzotriazole-1-base-N then in batches, N, N ', and N '-tetramethyl-urea hexafluorophosphate (48.6g, 128.15mmol).The solution that obtains stirred under room temperature, nitrogen atmosphere spend the night, be poured into then in the water (2.5L).Mixture is divided into two batches, every batch with ethyl acetate (2 *～750ml) extraction.The extract that clean to merge with salt solution (～500ml/ criticizes), and by dried over mgso, filter also evaporation obtains amber jelly (54g), with it with quick silicon-dioxide chromatogram purification, with pure EtOAc isocratic elution.Pure fraction is evaporated to drying, obtains viscous foam, by a small amount of EtOAc (～50ml) in mild heat, make its dissolving.Concentrate this solution slightly by revolving steaming then, and leave standstill.With solid pulp in ethyl acetate of gained～7 hours, standing over night then.Suction filtration is collected solid and 55 ℃ of following vacuum-drying a few hours, is obtained product then, is white solid (20.1g).This material demand is further purified, therefore this material is placed diethyl ether (～75-100ml), and stirred 2 hours.And then suction filtration collects solid and dry, and the material with independent placement merges then, and ethyl acetate (～60ml) in pulp a few hours.Mixture is left standstill a weekend, and suction filtration is collected solid then, cleans with more ethyl acetate, and 55 ℃ of vacuum-dryings to constant weight, obtain 4-(3-(4-oxyethyl group piperidines-1-carbonyl)-4-luorobenzyl) phthalazines-1 (2H)-ketone (2f) (21.70g, 58.4%), be white solid.

Embodiment 48

Restraining effect

For the restraining effect of assessing compound, use following assay method to determine IC ₅₀Value.

Will be from Hela nucleus extract isolating Mammals PARP is with Z damping fluid (25mMHepes (Sigma); 12.5mM MgCl ₂(Sigma); 50mM KCl (Sigma); 1mM DTT (Sigma); 10% glycerine (Sigma) 0.001%NP-40 (Sigma); PH7.4) at 96 hole FlashPlates (trade mark) (NEN, the described inhibitor of UK) the middle cultivation, and adding different concns.All compounds are diluted in DMSO, make final analysis concentration between 10 to 0.01 μ M, the ultimate density of DMSO is every hole 1%.Total mensuration volume in each hole is 40 μ l.

30 ℃ down cultivate 10 minutes after, add 10 μ l contain NAD (5 μ M), ³The reaction mixture initiation reaction of H-NAD and 30mer double-stranded DNA-oligomer.The specified positive and negative reaction hole and compound hole (the unknown) combination are handled to calculate enzymic activity %.With plate jolting 2 minutes, cultivated 45 minutes down then at 30 ℃.

After the cultivation, in each hole, add the acetate termination reaction of 50 μ l 30%.Then with plate jolting at room temperature 1 hour.

Plate is transferred to TopCount NXT (trade mark) (Packard, UK) enterprising line flicker counting.Record value is the counting (cpm) to the per minute of each hole counting after 30 seconds.

Use following formula to calculate the enzymic activity % of every kind of compound then:

Calculate IC ₅₀Value (concentration when suppressing 50% enzymic activity), it is measured in the different concns scope, is reduced to 0.001 μ M from 10 μ M usually.With this IC ₅₀Value is worth the compound efficacy that increases to differentiate as a comparison.

	IC ₅₀(nM)
		2a	7
2b	5
		2c	4
2d	4
		2e	4
2f	3
		2g	4
2h	4
		2i	4
2j	5
		4a	13
4b	5
		4c	16
6a	4
		6b	7
6c	16
		6d	7
6e	10

6f	8
		7	8
8a	3
		8b	2
8c	5
		8d	4
8e	5
		8f	2
9	7
		10	3
11	4
		12	7
15	10
		16	6
19	5
		20	5
24	6
		25	7
26a	4
		26b	10
26c	3

26d	6
		26e	6
26f	5
		26g	7
26h	3
		26i	2
26j	5
		26k	1
26l	5
		26m	7
29	3
		35a	479
36	4
		37	5
39	5
		40	6
46	11
		47a	10
47b	30
		52a	152
52b	55

54	18
		56	14
58	19
		60	26
63	69
		65	40
68	24
		69	8
70	6
		71	9
72a	664
		72b	677
72c	1739
		72d	1028
73	5
		74	8
77	8
		80	11
83	14
		86	18
89	7

90	9
		93	8
98	135
		99	11
104a	9
		104b	7
107	9
		110	22
113	8
		116	46
119a	21
		119b	13
126a	6
		126b	6
129a	4
		129b	23

Synergistic factor

Synergistic factor (the PF of compound ₅₀) be calculated as the IC of control cells growth ₅₀IC divided by cell growth behind the adding PARP inhibitor ₅₀Ratio.The growth-inhibiting curve of the cell of cellular control unit and compound treatment all has alkylating agent methyl mesylate (MMS) to exist down.Using fixed concentration is 0.2 micromolar test-compound.The concentration range of MMS is 0-10 μ g/ml.

Use sulphonyl rhodamine B (SRB) assay method (Skehan, P., Deng, (1990) New colorimetriccytotoxicity assay for anticancer-drug screening.J.Natl.Cancer Inst.82 1107-1112.) estimates the cell growth.The volume of 2,000 HeLa cells with 100 μ l is inoculated in each hole of flat 96 hole microtiter plates, cultivated 6 hours down at 37 ℃.Use independent nutrient solution or contain ultimate density and change cell as the nutrient solution of the PARP inhibitor of 30nM or 200nM.Make cell regeneration long 1 hour, then the MMS (being generally 0,1,2,3,5,7 and 10 μ g/ml) of a series of concentration of adding in untreated cell or the cell handled with the PARP inhibitor.The cell that uses the PARP inhibitor to handle is used to estimate the growth-inhibiting effect of PARP inhibitor separately.

Cell was placed 16 hours again, changed nutrient solution then, and make cell 37 ℃ of regrowths 72 hours.Remove nutrient solution then, with ice-cold 10% (w/v) trichoroacetic acid(TCA) fixed cell of 100 μ l.Described plate was cultivated 20 minutes down at 4 ℃, washed with water then four times.1% acetic acid solution of using 100 μ l 0.4% (w/v) SRB then is with the cell dyeing in each hole 20 minutes, then with 1% acetate washing four times.Made described plate then at room temperature dry 2 hours.The 10mM Tris alkali dissolution of adding 100 μ l is colored the dyestuff in the cell in each hole.The described plate of jolting was at room temperature placed 30 minutes gently, then under 564nM on Microquant microtitration plate reader measuring light density.

Patent	PF ₅₀(30nM)
		2b	3.0
2c	1.9
		2d	2.1
2e	1.6
		2f	14.1
2g	4.3
		2h	2.9
2i	16.7
		2j	3.0
4a	1.2
		4b	2.2
4c	1.2
		6a	1.7
6b	1.5
		6c	1.2

6d	1.4
		6e	1.5
6f	1.8
		7	4.0
8a	1.6
		8b	23.6
8c	12.2
		8d	1.2
8e	1.4
		8f	4.1
9	1.3
		10	3.1
11	33.6
		12	2.1
15	4.4
		16	3.4
19	2.5
		20	2.0
24	1.5
		25	2.1
26a	3.7
		26b	1.5
26c	32.4

26d	4.0
		26e	2.7
26f	4.4
		26g	1.6
26h	12.7
		26i	8.0
26j	1.2
		26k	26.8
26l	1.2
		26m	2.4
29	25.9
		36	14.5
37	1.7
		39	1.4
40	2.2
		46	20.1
47a	1.8
		54	1.3
58	1.0
		60	1.0
63	1.0
		69	1.7
70	32.1

71	4.0
		73	6.2
74	1.5
		77	3.1
80	1.5
		83	1.7
89	1.9
		90	1.1
93	2.8
		99	1.0
104a	1.3
		104b	2.2
107	1.7
		113	5.8
119a	1.0
		119b	1.2
126a	11.0
		126b	7.4
129a	1.0

	PF ₅₀(200nM)
		2a	3

2b	15
		2c	7
2d	11
		2e	8
2f	25
		6a	4.2
6b	3.5
		6c	1.5
6d	3.5
		6e	1.9
6f	5.0
		36	24.3
37	10.6
		40	19.8
46	3.24
		70	19.5
71	8.9
		73	27.8
113	23.1

Embodiment 49

To be derived from the cell of the Hela clone that is called KBA1, non--P-P-glycoprotein expression clone that it is expressed high P-glycoprotein (ABC1a and ABC1b transhipment glycoprotein also are called as MDR1a and MDR1b) and is called the coupling of KB31 is with 1.00 * 10 of every hole 80 μ l ⁴Individual cell/ml, promptly 800 cells/well [DMEM, 10%FBS, PSG] are inoculated in the 96 hole tissue culturing plates, and it was adhered to 4 hours.A kind of known P-gp inhibitor verapamil (Verapamil) (making ultimate density is 20 μ M) or the solid support medium that after incubation period, in the different holes of cell plate, add the 200 μ M of every hole 10 μ l.This 96 orifice plate is placed incubator 1 hour, in the hole of containing verapamil and substratum contrast hole, add the PBS/1%DMSO carrier (contrast hole) of 10 μ l test-compounds (or known Etoposide substrate as a comparison) or 10 μ l then.In different concentration ranges, common 100 μ M-0.3 μ M detect test-compound.

Tissue Culture Plate was cultivated 5 days, used aforesaid sulphonyl rhodamine B (SRB) assay method to measure the growth of cell afterwards.Use test-compound under the condition that has or do not exist (contrast hole) verapamil calculates the P-gp substrate activity of every kind of compound to the cells growth activity of KBA1 cell.Calculate the dosage correction than (DMR) by KBA1, it is the IC that does not add verapamil for every kind of test-compound ₅₀Divided by the cell growth IC that adds verapamil ₅₀Ratio.The compound that is not the P-gp substrate has＜1.5 DMR, and shown usually＞1.5 DMR, more typically greater than 2 by the compound that P-gp actively effluxes.

Compound 2b has 1.3 DMR, and compound 2f has 1.0 DMR.

Embodiment 50

By " Leach, A., etc., J Med Chem (2006), 49 (23), 6672-6682 " in the method described measure the solvability of test-compound:

By compound being stirred 24 hours, measure solubility values in the phosphoric acid buffer of the pH7.4 of 0.1M at 25 ℃.By two times centrifugal, with upper solution and undissolved separating substances, the standard of the concentration known of reference in DMSO is analyzed in conjunction with mass spectra peak identification with coventional type HPLC-UV method and is quantized then.

Compound 2b has the solubleness of 1070 μ Mol, and compound 2f has the solubleness of 211 μ Mol.

Claims

1. the compound of a formula (I):

Wherein:

A and B represent the optional fused aromatic rings that replaces together;

R ^CBe selected from H, C _1-4Alkyl; And

R ^CAnd R ¹Form spiral shell-C with carbon atom and Sauerstoffatom that they connected _5-7Oxygen containing heterocyclic radical, described heterocyclic radical are optional replace or and C _5-7The aromatic ring condensed.

2. compound according to claim 1, wherein-A-B-only is made up of carboatomic ring atom.

3. compound according to claim 2, wherein-A-B-forms phenyl ring.

4. compound according to claim 3, wherein said phenyl ring is unsubstituted.

5. compound according to claim 3, wherein said phenyl ring contain and are selected from halogen, NH ₂And C _1-4One or two substituting group of alkoxyl group.

6. according to each described compound of claim 1-5, wherein Y is CH.

7. according to each described compound of claim 1-6, wherein X is CH or CF.

8. according to each described compound of claim 1-6, wherein X is N.

9. according to each described compound, wherein R of claim 1-8 ¹Be H.

10. according to each described compound, wherein R of claim 1-8 ¹Be saturated C _1-7Alkyl.

11. compound according to claim 10, wherein R ¹Be methyl or ethyl.

12. each described compound, wherein R according to claim 1-9 ¹Be C ₆Aryl.

13. compound according to claim 12, wherein said C ₆Aryl is selected from phenyl, pyridyl, pyridazinyl, pyrimidyl and pyrazinyl.

14. a pharmaceutical composition comprises each described compound and pharmaceutical acceptable carrier or the thinner of claim 1-13.

15. the purposes of each described compound in treatment human body and animal body according to claim 1-13.

16. the purposes of each described compound in the active medicine of preparation inhibition PARP according to claim 1-13.

17. according to the purposes of each described compound in the preparation medicine of claim 1-13, described medicine is used for the treatment of: vascular disease; Septic shock; Ischemia injury; Neurotoxicity; Hemorrhagic shock; Virus infection; Perhaps by suppressing the active disease of improving of PARP.

18. according to each described compound of claim 1-13 purposes in the preparation medicine, described medicine is as the auxiliary of cancer therapy or be used to strengthen the therapeutic action to tumour cell of ionizing radiation or chemotherapeutic.

19. be used for purposes in the medicine of individuality treatment cancer according to each described compound of claim 1-13 in preparation, wherein said cancer is a HR dependent DNA DSB reparation approach disappearance.

20. purposes according to claim 19, wherein said cancer comprise, and one or more abilities by HR DNA plerosis DSB for normal cell reduce or the cancer cells of forfeiture.

21. purposes according to claim 20, wherein said cancer cells have the phenotype of BRCA1 or BRCA2 disappearance.

22. purposes according to claim 21, wherein said cancer cells are BRCA1 or BRCA2 disappearance.

23. according to each described purposes of claim 19-22, wherein said individuality is a heterozygosis with regard to the sudden change of the gene of coding HR dependent DNA DSB reparation pathway component.

24. purposes according to claim 23, wherein said individuality are heterozygosis with regard to the sudden change among BRCA1 and/or the BRCA2.

25. according to each described purposes of claim 19-24, wherein said cancer is mammary cancer, ovarian cancer, carcinoma of the pancreas or prostate cancer.

26. according to each described purposes of claim 19-25, wherein said treatment also comprises uses ionizing radiation or chemotherapeutic.